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Clinical Studies
Abstracts are presented below for clinical
studies on Indian Kudzu.
Plant Phytonutrient Profile
1: Phytochem Anal. 2007 Jan;18(1):50/9.
Identification of isoflavone glycosides in Pueraria lobata cultures by tandem
mass spectrometry.
Prasain JK, Reppert A, Jones K, Moore DR 2nd, Barnes S, Lila MA.
Department of Pharmacology and Toxicology, University of Alabama at Birmingham,
Birmingham, AL 35294, USA. jprasain@uab.edu
Isoflavones in the methanolic extracts of kudzu (Pueraria lobata) callus,
suspension and root cultures were compared in order to develop an experimental
system in which puerarin (daidzein 8/C/glucoside) and other isoflavones could be
synthesised in vitro. Quantitative variation of puerarin and other known
isoflavones was estimated in kudzu culture extracts using HPLC/UV. The highest
and lowest amounts of puerarin (14.56 and 0.33 mg/g) were present in in vitro
root cultures and leaf tissue/derived callus cultures, respectively. A total of
48 isoflavone metabolites were detected in extracts of kudzu root cultures by
HPLC/MS/MS, and the structures of 33 of them were tentatively assigned. Amongst
these, 12 isoflavone C/glycosides were identified. Hydroxyderivatives of
puerarin in several isomeric forms were detected, some of which have not been
previously reported in kudzu root. The molecular weights, interpretation of
characteristic fragment ions obtained from HPLC/MS/MS and comparison with
reported data allowed the putative identification of the isoflavone metabolites.
Publication Types:
Research Support, N.I.H., Extramural
PMID: 17260698 [PubMed / indexed for MEDLINE]
2: Planta Med. 1998 Oct;64(7):620/7.
Identification of Isoflavones in the Roots of Pueraria lobata.
Rong H, Stevens JF, Deinzer ML, Cooman LD, Keukeleire DD.
University of Gent, Faculty of Pharmaceutical Sciences, Gent, Belgium.
The isoflavones of the roots of PUERARIA LOBATA (Willd.) Ohwi (Puerariae Radix)
were investigated by high/performance liquid chromatography (HPLC) coupled to
photodiode array (PDA) and to mass spectroscopy (MS) using atmospheric pressure
chemical ionization (APCI) or electrospray ionization (ESI) in combination with
collision/activated decomposition (CAD) (HPLC/APCI/CAD/MS or ESI/CAD/MS) for
identification of glycosides and HPLC/APCI/CAD/MS for identification of
aglycones. The major glycosides are derived from daidzein ( 9) and most are 8/
C/glycosides. 3'/Hydroxypuerarin/4'/ O/deoxyhexoside ( 2B) and 3'/methoxy/6''/
O/ D/xylosylpuerarin ( 6) were identified as new constituents. MS data were
obtained for puerarin/4'/ O/ D/glucoside ( 1), 3'/hydroxypuerarin ( 2A),
puerarin ( 3), 3'/methoxypuerarin ( 4), 6''/ O/ D/xylosylpuerarin ( 5), daidzin
( 7) and 3'/methoxydaidzin ( 8), which were previously characterized by NMR
analysis. Isoflavones identified in Puerariae Radix comprise 3'/methoxydaidzein
( 10), genistein ( 12), daidzein/7/ O/methyl ether ( 13A), 3'/methoxydaidzein/7/
O/methyl ether or 3'/methoxyformononetin ( 13B) and biochanin A ( 15), while
previous characterization of daidzein ( 9) and formononetin ( 14) was
substantiated by MS data. The structure of compound 11 could not be established
by MS techniques. The estrogenic activity was mainly located in the aglycone
fraction.
PMID: 17253303 [PubMed / in process]
3: J Reprod Dev. 2007 Jan 17; [Epub ahead of print]
Evaluation of the Estrogenic Activity of the Wild Pueraria mirifica by Vaginal
Cornification Assay.
Cherdshewasart W, Kitsamai Y, Malaivijitnond S.
Department of Biology, Faculty of Science, Chulalongkorn University.
The aim of this study was to evaluate the estrogenic activity of tuberous
samples of phytoestrogen/rich Pueraria mirifica collected from 25 of 76
provinces in Thailand by vaginal cornification assay. Tuberous powders were
prepared and administered to ovariectomized rats for 14 consecutive days at
dosages of 10, 100, and 1,000 mg/kg BW respectively, and were compared with a
daily treatment with 200 mug/100 g BW 17beta/estradiol (E2). Rats treated with
10 mg/kg BW Pueraria mirifica showed no vaginal cornification. Treatment with
100 mg/kg BW Pueraria mirifica from 13 out of 25 plant samples resulted in
development of vaginal cornification. The cell count percentages of the vaginal
smeared cells for the treatment with the 2 plant samples that exhibited the
fastest vaginal cornification revealed large variation in their estrogenic
activities. Treatment with 1,000 mg/kg BW Pueraria mirifica from all plant
samples produced vaginal cornification with the mean value for the period (day)
of first appearance of cornified cells being 4.08 days compared to 2 days with
200 mug/100g BW E2. The overall appearance period (day) of cornified cells
during the treatment and post/treatment period with 1,000 mg/kg BW per day
Pueraria mirifica was shorter than treatment with 200 mug/100 g BW E2. The
results demonstrate that the plant population shows differential estrogenic
activity as evaluated by vaginal cornification assay.
PMID: 17229996 [PubMed / as supplied by publisher]
4: J Reprod Dev. 2006 Dec 29; [Epub ahead of print]
Assessment of Urinary Gonadotropin and Steroid Hormone Profiles of Female
Cynomolgus Monkeys after Treatment with Pueraria mirifica.
Trisomboon H, Malaivijitnond S, Cherdshewasart W, Watanabe G, Taya K.
Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn
University.
This study investigated the changes in the urinary hormone levels of female
monkeys (Macaca fascicularis) after single/dose and long/term treatments with
Pueraria mirifica (PM). The monkeys were separated into 3 groups (n=3) and
orally treated with 10, 100, or 1,000 mg of PM in each group. Two series of
experiments were performed. In the first series of experiments, the monkeys were
orally treated with a single dose of PM. The experimental schedule was divided
into a one menstrual cycle pretreatment period and a two menstrual cycle
post/treatment period. In the second series of experiments, the monkeys were
orally treated daily with PM for 90 days. The experiment schedule was divided
into a one menstrual cycle pretreatment period, a three menstrual cycle
treatment period, and a two menstrual cycle post/treatment period. Urinary
samples were collected daily and assayed for the FSH, LH, estradiol, and
progesterone levels. The results showed that there were no changes in the FSH,
LH, estradiol, and progesterone levels after treatment with a single dose of 10,
100, or 1,000 mg of PM or after daily treatment with 10 mg of PM for 90 days
compared with the levels observed during the pretreatment period. Daily
treatment with 100 mg and 1,000 mg of PM for 90 days only produced a clear
reduction in the urinary FSH levels. This suggests that changes of urinary FSH
levels can be considered an indicator for study of estrogenic effects on
hormonal levels in female monkeys.
PMID: 17202751 [PubMed / as supplied by publisher]
5: Biol Pharm Bull. 2007 Jan;30(1):173/5.
Molecular authentication of Radix Puerariae Lobatae and Radix Puerariae
Thomsonii by ITS and 5S rRNA spacer sequencing.
Sun Y, Shaw PC, Fung KP.
Department of Biochemistry and Institute of Chinese Medicine, The Chinese
University of Hong Kong, NT.
In the present study, we examined nuclear DNA sequences in an attempt to reveal
the relationships between Pueraria lobata (Willd). Ohwi, P. thomsonii Benth.,
and P. montana (Lour.) Merr. We found that internal transcribed spacer (ITS)
sequences of nuclear ribosomal DNA are highly divergent in P. lobata and P.
thomsonii, and four types of ITS with different length are found in the two
species. On the other hand, DNA sequences of 5S rRNA gene spacer are highly
conserved across multiple copies in P. lobata and P. thomsonii, they could be
used to identify P. lobata, P. thomsonii, and P. montana of this complex, and
may serve as a useful tool in medical authentication of Radix Puerariae Lobatae
and Radix Puerariae Thomsonii.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17202681 [PubMed / in process]
6: Endocrine. 2006 Aug;30(1):93/101.
Effects of Pueraria mirifica, an herb containing phytoestrogens, on reproductive
organs and fertility of adult male mice.
Jaroenporn S, Malaivijitnond S, Wattanasirmkit K, Trisomboon H, Watanabe G, Taya
K, Cherdshewasart W.
Interdepartment of Physiology, Faculty of Graduate School, Chulalongkorn
University, Bangkok 10330, Thailand.
The effects of Pueraria mirifica (PM) on reproductive organs and fertility of
adult male mice were investigated. Male mice were divided into four groups (10
mice/group). Groups 1/3 were orally treated with PM at doses of 0 (PM/0), 10
(PM/10), and 100 (PM/100) mg/kg BW/d in 0.2 mL distilled water, and group 4 was
subcutaneously injected with 200 microg/kg BW/d of synthetic estrogen
diesthylstilbestol (DES). The treatment schedule was separated into two periods:
treatment and posttreatment (8 wk for each period). The PM/10 and PM/100
treatments had no effect on testicular weight, sperm number, and serum LH, FSH,
and testosterone levels. Only the PM/100 treatment reduced weights of epididymes
and seminal vesicle and the sperm motility and viability. Histopathological
examination demonstrated that testis, epididymis, and seminal vesicle were
normal in all doses of PM treatment. PM/treated males showed no alterations in
mating efficiency and on causing pregnancy of their female partners. DES
injection impaired all those parameters. Offspring fathered by the PM/ and
DES/treated males exhibited neither malformations nor change of body weight
gains, and the reproductive organ weights of 50/d old pups were in the normal
range. The present data clearly demonstrate that a long/term treatment of PM at
doses 10 and 100 mg/kg BW/d, via oral route, does not alter a male fertility and
a hypothalamus/ pituitary/testis axis. Although PM/100 can cause some moderate
impairment, no persistent effects were observed. Most of PM/treated mice
increased the mating efficiency after stop treatment.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17185797 [PubMed / indexed for MEDLINE]
7: Indian J Exp Biol. 2006 Dec;44(12):1012/7.
Isoflavonoids production in callus culture of Pueraria tuberosa, the Indian
kudzu.
Vaishnav K, Goyal S, Ramawat KG.
Laboratory of Biomolecular Technology, Department of Botany, M L Sukhadia
University, Udaipur 313 001, India.
Isoflavonoid contents of different plant parts and callus tissues of the Indian
Kudzu, Pueraria tuberosa (Roxb.ex.Willd.) DC are presented. The initial cultures
were slow growing, associated with browning of the tissues. The production of
four isoflavonoids (puerarin, genistin, genistein and daidzein) in the callus
cultures of P. tuberosa was studied by manipulating the plant growth regulators
and sucrose concentration in the medium. Organogenesis was not recorded in
callus on any of these treatments. Tuber and stem accumulated puerarin, a
glycoside of daidzein, at high amounts, 0.65% and 0.054% respectively. However,
the daidzein content of the callus tissues grown on Murashige and Skoog medium
containing BA (20.9 microM) and sucrose (60 gl(/1)) was significantly higher
(0.056%) than in vivo plant material (0.02%) and other comparable culture
systems like Genista and Pueraria lobata.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17176676 [PubMed / indexed for MEDLINE]
8: Zhongguo Zhong Yao Za Zhi. 2006 Oct;31(19):1580/3.
[Effect of several physiochemical factors on cell growth and isoflavone
accumulation of Pueraria lobata cell suspension culture]
[Article in Chinese]
Fang CB, Li HQ, Wan XC, Jiang CJ.
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture,
Anhui Agricultural University, Hefei 230036, China.
OBJECTIVE: To illustrate the effects of several physiochemical factors on cell
growth and isoflavone accumulation of Pueraria lobata cell suspension cultures.
METHOD: High performance liquid chromatography and plant tissue culture were
applied. RESULT AND CONCLUSION: Cell growth and isoflavone accumulation were
significantly stimulated in P. lobata cell suspension cultures by the increase
of the sucrose concentration. Maintaining the pH value at the range over 5. 4 to
5. 8 was most suitable for isoflavone accumulation in P. lobata cell suspension
cultures. Cell dried weight and isoflavone accumulation decreased sharply with
the increase of the treated concentration of active carbon, while XAD/4
significantly stimulated cell growth and isoflavone accumulation.
Publication Types:
English Abstract
PMID: 17165578 [PubMed / in process]
9: Mol Cell Biochem. 2006 Dec 6; [Epub ahead of print]
A Lupinoside prevented fatty acid induced inhibition of insulin sensitivity in
3T3 L1 adipocytes.
Dey D, Pal BC, Biswas T, Roy SS, Bandyopadhyay A, Mandal SK, Giri BB,
Bhattacharya S.
Indian Institute of Chemical Biology, 4 Raja S.C.Mullick Road, Kolkata, 700032,
India.
The decrease in insulin sensitivity to target tissues or insulin resistance
leads to type 2 diabetes mellitus, an insidious disease threatening global
health. Numerous evidences made free fatty acids (FFAs) responsible for insulin
resistance and type 2 diabetes. We demonstrate here that the damage of insulin
acitivity by a free fatty acid, palmitate could be prevented by a lupinoside. An
incubation of 3T3 L1 adipocytes with a FFA i.e. palmitate inhibited insulin
stimulated uptake of (3)H/2 deoxyglucose (2 DOG) significantly. Addition of a
lupinoside purified from Pueraria tuberosa, lupinoside PA(4) (LPA(4)) strongly
prevented this inhibition. We then examined insulin signaling pathway where
palmitate significantly inhibited insulin stimulated phosphorylation of Insulin
receptor tyrosine kinase, IRS 1and PI3 kinase, PDK1 and Akt/PKB. LPA(4) rescued
this inhibition of signaling molecule by palmitate. Insulin mediated
translocation of Glut4, the glucose transporter in insulin target cells, was
effectively blocked by palmitate while, LPA(4) waived this block. Administration
of LPA(4) to nutritionally induced diabetic rats significantly reduced the
increase in plasma glucose. All these indicate LPA(4) to be a potentially
therapeutic agent for insulin resistance and type 2 diabetes.
PMID: 17149545 [PubMed / as supplied by publisher]
10: Biol Pharm Bull. 2006 Dec;29(12):2432/5.
Intestinal bacteria activate estrogenic effect of main constituents puerarin and
daidzin of Pueraria thunbergiana.
Park EK, Shin J, Bae EA, Lee YC, Kim DH.
College of Pharmacy, Kyung Hee University, Seoul, Korea.
To understand the relationship between the metabolites and estrogenic activity
of the main isoflavones puerarin and daidzin of the rhizome of Pueraria
thunbergiana (PT, family Leguminosae), PT and its isoflavones were transformed
by human intestinal bacteria and their estrogenic effects were investigated. All
human fecal specimens hydrolyzed puerarin and daidzin to daidzein, but their
hydrolyzing activities varied depending on the individuals. All intestinal
bacteria isolated from human also hydrolyzed daidzin to daidzein, but a few
bacteria transformed puerarin to daidzein. When the estrogenic effect of PT,
puerarin and daidzin was compared with those of their metabolites, the
metabolites more potently increased proliferation of MCF/7 cells than PT,
puerarin and daidzin. The metabolite daidzein also potently increased
estrogen/response c/fos mRNA and PR protein expressions. These findings suggest
that intestinal bacteria, which can hydrolyze puerarin and/or daidzin, may
activate a potent estrogenic activity of PT.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17142977 [PubMed / indexed for MEDLINE]
11: Arch Pharm Res. 2006 Oct;29(10):821/5.
Constituents of the roots of Pueraria lobata inhibit formation of advanced
glycation end products (AGEs).
Kim JM, Lee YM, Lee GY, Jang DS, Bae KH, Kim JS.
Department of Herbal Pharmaceutical Development, Korea Institute of Oriental
Medicine, Daejeon 305/811, Korea.
Two isoflavone C/glucosides, puerarin (1) and PG/3 (2), a but/2/enolide,
(+//)/puerol B (3), two isoflavone O/glucosides, daidzin (4) and genistin (5),
and three pterocarpans, (/)/medicarpin (6), (/)/glycinol (7) and (/)/tuberosin
(8), were isolated from a MeOH extract of the roots of Pueraria lobata, using an
in vitro bioassay based on the inhibition of the formation of advanced glycation
end products (AGEs) to monitor chromatographic fractionation. The structures of
1/8 were determined by spectroscopic data interpretation, particularly by 1D/
and 2D/NMR studies, and by comparison of these data with values in the
literature. All of the isolates (1/8) were evaluated for their inhibitory
activity on AGEs formation in vitro. Of these, puerarin (1), PG/3 (2), and
(+//)/puerol B (3) exhibited more potent inhibitory activity than the positive
control aminoguanidine.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17121174 [PubMed / indexed for MEDLINE]
12: Maturitas. 2007 Mar 20;56(3):322/31. Epub 2006 Nov 13.
Pueraria mirifica, a phytoestrogen/rich herb, prevents bone loss in
orchidectomized rats.
Urasopon N, Hamada Y, Asaoka K, Cherdshewasart W, Malaivijitnond S.
Biological Science Ph.D. Program, Faculty of Science, Chulalongkorn University,
Bangkok 10330, Thailand; Primate Research Unit, Department of Biology, Faculty
of Science, Chulalongkorn University, Bangkok 10330, Thailand.
OBJECTIVE: Estrogens and estrogen/like substances have been reported to play an
important role in male bone homeostasis and to prevent bone loss. Pueraria
mirifica (Leguminosae), a Thai herbal plant, containing a high amount of
phytoestrogens was a choice of interest for this study. We examined the effects
of crude P. mirifica on bone loss and influences on reproductive organs in male
rats. METHODS: Using fully mature and orchidectomized (ORX) rats, the effects of
0, 10, 100 and 1000mg/kgB.W./day of P. mirifica and 0.1mg/kgB.W./day of 17
alpha/ethinylestradiol (a positive control) were evaluated on bone mineral
density (BMD) and bone mineral content (BMC) measured with a peripheral
Quantitative Computerized Tomography (pQCT) densitometry. RESULTS: Bone loss in
trabecular and cortical bones of the various sites of axial bone (fourth lumbar
vertebral body) and long bones (tibia and femur) after ORX was dose/dependently
prevented by P. mirifica. The effects were specific on bone types and sites. The
weights of the accessory sex organs, seminal vesicle and ventral prostrate
gland, which significantly decreased after 3/month of ORX, were not altered by
P. mirifica. CONCLUSION: The results suggest that P. mirifica treatment may be
useful to prevent an osteoporosis in elderly hypogonadism subjects without
influences on reproductive organs.
PMID: 17101247 [PubMed / in process]
13: Zhong Yao Cai. 2006 Aug;29(8):816/8.
[Studies on the effect of extracts of several Chinese herbal medicines and other
medicines on alcohol dehydrogenase activity]
[Article in Chinese]
Li WZ, Lu J, Sun XY, Wang SL, Ni YM, Zhu WS.
College of Life Science and Technology of Tongji University, Shanghai 200092,
China. lwz@mail.tongji.edu.cn
OBJECTIVE: To study the effects of water and alcohol extracts of several Chinese
herbal medicines and other medicines on alcohol dehydrogenase activity in order
to provide enzymology basis on new medicine. METHODS: Water or alcohol extracts
of Chinese herbal medicine and other medicine were tested on the effects of
alcohol dehydrogenase activity by Valle and Hoch method. RESULTS: Among them, 8
were found to have the effect of activation on alcohol dehydrogenase. They were
water extracts of Amomum kravanh and Pueraria flowers, the alcohol extracts of
Pueraria flowers, compound hepatcare Chinese medicine and compound Pueraria
medicine, L/cysteine, notoginseng saponin. Others had inhibiting action.
CONCLUSION: To decrease alcohol concentration in the body through activating the
activity of ADH may be one of the mechanisms for some traditional Chinese herbal
medicine in neutralizing the effect of alcohol drink.
Publication Types:
English Abstract
PMID: 17076243 [PubMed / in process]
14: Wei Sheng Wu Xue Bao. 2006 Aug;46(4):653/6.
[Primary study on acid tolerance mechanism of a wild aciduric Rhizobium strain
isolated from Pueraria lobata]
[Article in Chinese]
Gu JP, Zhang L, Wei SQ, Zhang Q, Fang DH.
College of Resource and Environment, Southwest University, Chongqing 400716,
China. gwping80@163.com
An aciduric Rhizobium strain, named as PR389, was isolated from the nodule of
wild Pueraria lobata which grew in yellow soil (pH 4.6) on the Jin/yun Mountain
in Bei/bei, Chong/qing city. The isolated strain, which could grow under pH 4.6
distinct from the optimal pH 6.5/7.5 for rhizobium, showed some typical features
of aciduric rhizobium. This was also confirmed by the proton flux assay.
Compared to the acid/sensitive Rhizobium strain PR21, the cell membrane of PR389
could hold back excessive H entering cell. This feature can protect PR389 from
harm of acid. In the test of acid tolerance, the aciduric ability of strain
PR389 under low acidic (pH 3.8) was restrained by antibiotic chloramphenicol. It
was speculated that special proteins in the cells of PR389 could be induced and
synthesized in acidic environment.
Publication Types:
English Abstract
PMID: 17037073 [PubMed / in process]
15: Eur J Pharmacol. 2006 Dec 15;552(1/3):105/11. Epub 2006 Sep 14.
Puerarin, an isoflavonoid derived from Radix puerariae, potentiates
endothelium/independent relaxation via the cyclic AMP pathway in porcine
coronary artery.
Yeung DK, Leung SW, Xu YC, Vanhoutte PM, Man RY.
Department of Pharmacology, 2/F Faculty of Medicine Building, University of Hong
Kong, 21 Sassoon Road, Pokfulam, Hong Kong, People's Republic of China.
Puerarin, an isoflavonoid derived from the Chinese medicinal herb Radix
puerariae, has been suggested to be useful in the management of various
cardiovascular disorders. The present study examined the effect of acute
exposure (30 min) to puerarin on vascular relaxation. Rings from porcine
coronary artery of either sex were used. The highest concentration of puerarin
(100 microM) produced a small but statistically significant relaxation of
U46619/contracted rings. Vascular relaxations were also studied in the presence
of lower concentrations of puerarin (0.1, 1 and 10 microM) which had no direct
relaxation effect. Puerarin enhanced vasorelaxation to endothelium/independent
relaxing agents, sodium nitroprusside and cromakalim. However, puerarin had no
effect on vasorelaxation induced by endothelium/dependent relaxing agents,
bradykinin and calcium ionophore A23187. The potentiating action of puerarin (10
microM) on sodium nitroprusside/mediated relaxation was not affected by the
nitric oxide synthase inhibitor, N(omega)/nitro/L/arginine methyl ester (L/NAME;
300 microM), or by the disruption of the endothelium with Triton X/100. The
effect of puerarin was reversible following a washout period. The potentiating
effects were comparable with the 3'/5'/cyclic adenosine monophosphate (cyclic
AMP) analogues, 8/bromoadenosine/3'/5'/cyclic monophosphate (8/Br/cyclic AMP; 10
muM) and Sp/isomer [S nomenclature refers to phosphorus] of adenosine/3',
5'/cyclic monophosphorothioate (Sp/cyclic AMPS; 3 microM), but not the
3'/5'/cyclic guanosine monophosphate (cyclic GMP) analogue,
8/bromoguanosine/3'/5'/cyclic monophosphate (8/Br/cyclic GMP; 3 microM). The
cyclic AMP antagonist, Rp/isomer [R nomenclature refers to phosphorus] of
8/bromoadenosine/3', 5'/cyclic monophosphorothioate (Rp/8/Br/cyclic AMPS; 10
microM), but not cyclic GMP antagonist, Rp/isomer of 8/bromoguanosine/3',
5'/cyclic monophosphorothioate (Rp/8/Br/cyclic GMPS; 10 microM), reversed the
effects of puerarin (10 microM) on the enhancement of vasorelaxation to sodium
nitroprusside. Our results demonstrated that puerarin enhanced sodium
nitroprusside/induced relaxation, possibly via the cyclic AMP/dependent pathway.
Publication Types:
In Vitro
Research Support, Non/U.S. Gov't
PMID: 17027964 [PubMed / indexed for MEDLINE]
16: Sci China C Life Sci. 2006 Aug;49(4):379/89.
Nitric oxide mediates the fungal elicitor/induced puerarin biosynthesis in
Pueraria thomsonii Benth. suspension cells through a salicylic acid
(SA)/dependent and a jasmonic acid (JA)/dependent signal pathway.
Xu M, Dong J, Zhu M.
Department of Biotechnology, Zhejiang Gongshang University, Hangzhou 310035,
China. maojunxu@163.com
Nitric oxide (NO) has emerged as a key signaling molecule in plant secondary
metabolite biosynthesis recently. In order to investigate the molecular basis of
NO signaling in elicitor/induced secondary metabolite biosynthesis of plant
cells, we determined the contents of NO, salicylic acid (SA), jasmonic acid
(JA), and puerarin in Pueraria thomsonii Benth. suspension cells treated with
the elicitors prepared from cell walls of Penicillium citrinum. The results
showed that the fungal elicitor induced NO burst, SA accumulation and puerarin
production of P. thomsonii Benth. cells. The elicitor/induced SA accumulation
and puerarin production was suppressed by nitric oxide specific scavenger cPITO,
indicating that NO was essential for elicitor/induced SA and puerarin
biosynthesis in P. thomsonii Benth. cells. In transgenic NahG P. thomsonii
Benth. cells, the fungal elicitor also induced puerarin biosynthesis, NO burst,
and JA accumulation, though the SA biosynthesis was impaired. The
elicitor/induced JA accumulation in transgenic cells was blocked by cPITO, which
suggested that JA acted downstream of NO and its biosynthesis was controlled by
NO. External application of NO via its donor sodium nitroprusside (SNP) enhanced
puerarin biosynthesis in transgenic NahG P. thomsonii Benth. cells, and the
NO/triggered puerarin biosynthesis was suppressed by JA inhibitors IBU and NDGA,
which indicated that NO induced puerarin production through a JA/dependent
signal pathway in the transgenic cells. Exogenous application of SA suppressed
the elicitor/induced JA biosynthesis and reversed the inhibition of IBU and NDGA
on elicitor/induced puerarin accumulation in transgenic cells, which indicated
that SA inhibited JA biosynthesis in the cells and that SA might be used as a
substitute for JA to mediate the elicitor/ and NO/induced puerarin biosynthesis.
It was, therefore, concluded that NO might mediate the elicitor/induced puerarin
biosynthesis through SA/ and JA/dependent signal pathways in wildtype P.
thomsonii Benth. cells and transgenic NahG cells respectively.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16989284 [PubMed / indexed for MEDLINE]
17: Addict Biol. 2006 Sep;11(3/4):356/73.
Modulation of high alcohol drinking in the inbred Fawn/Hooded (FH/Wjd) rat
strain: implications for treatment.
Overstreet DH, Rezvani AH, Cowen M, Chen F, Lawrence AJ.
Bowles Center for Alcohol Studies & Department of Psychiatry, University of
North Carolina at Chapel Hill, USA. dhover@med.unc.edu
The Fawn/Hooded rat (FH/Wjd) is an inbred alcohol/preferring rat strain, unlike
most of the other strains that were selectively bred for high alcohol intake and
preference. It was chosen for study some 16 years ago because of a reported
mutation that disrupted platelet serotonin function. Although the FH/Wjd rat has
high alcohol intake (>5 g/kg/day) and preference (>65%), interbreeding with an
alcohol/non/preferring inbred strain suggested that these measures are unrelated
to the serotonin abnormality. Similarly, the exaggerated immobility of the
FH/Wjd rats in the forced swim test did not correlate with the high alcohol
intake. Many compounds have been tested in the FH/Wjd rats after both acute and
chronic treatment and a substantial number of them have proved effective.
However, as the case with opiate antagonists, tolerance to the effects of the
drug can develop. An up/regulation of opioid receptors accompanied the chronic
treatment and this mechanism may account for the development of tolerance.
Tolerance to opiate antagonists has also been demonstrated in two of the
selectively bred alcohol/preferring rat lines, but it is unknown whether this
process may contribute to the relapses seen in individuals being treated with
naltrexone. Other drugs that reliably decrease alcohol intake in the FH/Wjd rats
include the 5/hydroxytryptamine/2A receptor antagonist, amperozide, the mGlu5
receptor antagonist 3/[(2/methyl/1,3/thiazol/4/yl)ethynyl]pyridine (MTEP) and
herbal derivatives such as ibogaine, St. John's wort and kudzu extract. Thus,
studies in the FH/Wjd rat have led to the discovery of a wide variety of targets
for the development of novel agents to treat alcoholism. The fact that several
of these drugs were shown to reduce alcohol intake in some of the selectively
bred alcohol/preferring rat lines and/or alcohol/preferring vervet monkeys
increases our confidence that they are good candidates for further development.
Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non/U.S. Gov't
Review
PMID: 16961764 [PubMed / indexed for MEDLINE]
18: Appl Microbiol Biotechnol. 2007 Jan;73(5):1143/9. Epub 2006 Sep 8.
One/pot synthesis of genistein from tyrosine by coincubation of genetically
engineered Escherichia coli and Saccharomyces cerevisiae cells.
Katsuyama Y, Miyahisa I, Funa N, Horinouchi S.
Department of Biotechnology, Graduate School of Agriculture and Life Sciences,
The University of Tokyo, Bunkyo/ku, Tokyo, 113/8657, Japan.
For production of genistein from N/acetylcysteamine/attached p/coumarate
(p/coumaroyl/NAC) supplemented to the medium, a chalcone synthase (CHS) gene
from Glycyrrhiza echinata, a chalcone isomerase (CHI) gene from Pueraria lobata,
and an isoflavone synthase (IFS) gene from G. echinata were placed under the
control of the galactose/inducible GAL promoters in pESC vector and were
introduced in Saccharomyces cerevisiae. When the recombinant yeast cells (0.5 g
wet weight) were used as "enzyme bags" and incubated at 30 degrees C for 48 h in
100 ml of the buffer containing galactose and 1 mM (265 mg/l) p/coumaroyl/NAC,
ca. 340 microg genistein/l was produced. Another system consisting of two enzyme
bags was also generated for the purpose of production of genistein from
tyrosine. One enzyme bag was an Escherichia coli cell containing a phenylalanine
ammonia/lyase gene from a yeast, a 4/coumarate/cinnamate:CoA ligase gene from
the actinomycete Streptomyces coelicolor A3(2), the CHS gene, and the CHI gene,
in addition to the acetyl/CoA carboxylase gene from Corynebacterium glutamicum,
all of which were under the control of the
isopropyl/beta/D/thiogalactopyranoside/inducible T7 promoter, and thus producing
(S)/naringenin from tyrosine. The other enzyme bag was a S. cerevisiae cell
containing the IFS gene. Coincubation of the E. coli cells (0.5 g wet weight)
and S. cerevisiae cells (0.5 g wet weight) at 26 degrees C for 60 h in 20 ml of
the buffer containing 3 mM (543 mg/l) tyrosine as the starting substrate yielded
ca. 6 mg genistein/l.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16960736 [PubMed / in process]
19: Chem Pharm Bull (Tokyo). 2006 Sep;54(9):1315/7.
Puerariafuran, a new inhibitor of advanced glycation end products (AGEs)
isolated from the roots of Pueraria lobata.
Jang DS, Kim JM, Lee YM, Kim YS, Kim JH, Kim JS.
Department of Herbal Pharmaceutical Development, Korea Institute of Oriental
Medicine, 461/24 Jeonmin/dong, Yuseong/gu, Daejon 305/811, Korea.
A new 2/arylbenzofuran, puerariafuran (1), as well as three known compounds,
coumestrol (2), daidzein (3), and genistein (4), were isolated from a MeOH
extract of the roots of Pueraria lobata as active constituents, using an in
vitro bioassay based on the inhibition of advanced glycation end products (AGEs)
to monitor chromatographic fractionation. The structure of 1 was determined by
spectroscopic data interpretation, particularly by extensive 1D and 2D NMR
studies. All the isolates (1/4) were evaluated for the inhibitory activity on
AGEs formation in vitro.
Publication Types:
In Vitro
Research Support, Non/U.S. Gov't
PMID: 16946543 [PubMed / indexed for MEDLINE]
20: J Pharm Biomed Anal. 2007 Jan 17;43(2):428/34. Epub 2006 Aug 22.
Major isoflavonoid contents of the phytoestrogen rich/herb Pueraria mirifica in
comparison with Pueraria lobata.
Cherdshewasart W, Subtang S, Dahlan W.
Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai
Road, Patumwan, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th
Pueraria mirifica tubers collected from 28 out of 76 provinces of Thailand and
Pueraria lobata tubers collected from Guangzhou province, China were submitted
to HPLC analysis with the established gradient system comprising 1.5% acetic
acid and acetonitrile. Five major isoflavonoids, including puerarin, daidzin,
genistin, daidzein and genistein, were adopted as authentic standards. P.
mirifica tubers showed intra/ as well as inter/provincial differences in
isoflavonoid and total isoflavonoid contents. The difference in both cases
should be mostly influenced by genetic and environmental factors. In comparison
with P. lobata, P. mirifica population exhibited differences only with a lower
amount of daidzein.
PMID: 16930918 [PubMed / in process]
21: J Altern Complement Med. 2006 Jul/Aug;12(6):543/8.
Pharmacokinetic profile of the isoflavone puerarin after acute and repeated
administration of a novel kudzu extract to human volunteers.
Penetar DM, Teter CJ, Ma Z, Tracy M, Lee DY, Lukas SE.
Behavioral Psychopharmacology Research Laboratory, McLean Hospital, Belmont, MA
02478, USA. dpenetar@mclean.harvard.edu
OBJECTIVES: This study was undertaken to assess the pharmacokinetic profile of
puerarin, the major isoflavone found in a kudzu (Pueraria lobata) extract after
acute and repeated administration. METHODS: Participants were given either
single or repeated doses of kudzu extract, and blood samples were collected for
either 8 or 72 hours for subsequent pharmacokinetic analyses of puerarin.
RESULTS: Using WinNonlin pharmacokinetic data analysis software, puerarin was
found to be rapidly absorbed via the oral route, reach peak levels at 2 hours,
and have a half/life of approximately 4.3 hours. The elimination half/life was
not significantly altered after repeated administration. CONCLUSIONS: A
formulation of kudzu extract delivers a large amount of the principal isoflavone
in a rapid manner. The elimination rate constants and the mono/exponential
decline in blood levels suggest that a one compartment model adequately explains
how puerarin is handled by the body. Three times a day dosing is recommended as
accumulation will not occur, and plasma levels remain at levels that are
biologically active, even 8 hours after the last steady/state dose.
Publication Types:
Clinical Trial
Research Support, N.I.H., Extramural
PMID: 16884345 [PubMed / indexed for MEDLINE]
22: J Med Assoc Thai. 2005 Jun;88 Suppl 1:S21/9.
Effects of Pueraria mirifica on vascular function of ovariectomized rabbits.
Wattanapitayakul SK, Chularojmontri L, Srichirat S.
Department of Pharmacology, Faculty of Medicine, Srinakharinwirot University,
Sukhumvit 23, Wattana, Bangkok 10110, Thailand. suvara@swu.ac.th
Estrogen stimulates endothelial nitric oxide (NO) production and attenuates
endothelial dysfunction in ischemia/repurfusion and menopause. Recent studies
have shown that phytoestrogens from dietary sources improve endothelial function
and reduce cardiovascular risks. The Thai medicinal plant Pueraria mirifica (PM)
contains many potent phytoestrogens including miroestrol and deoxymiroestrol but
no study on vascular function has been established. Ground powder of PM was
orally given to ovariectomized White New Zealand rabbits (OVX + PM group) (n =
4) weighing 3.2/4.0 kg at the dose of 100 mg/kg for 90 days. Saline/treated
ovariectomized rabbits were assigned as a control group (OVX group) (n = 5). At
the end of treatment thoracic aorta was isolated for functional evaluation.
Maximal relaxant response to acetylcholine (ACh) was significantly increased
(24%) with 3.5/fold decrease in EC50 while no change in relaxant response to
sodium nitroprusside was observed Minimal and maximal responses to
17beta/estradiol (E2) were increased in the OVX + PM group and L/NAME (100 mM)
attenuated Emax of E2. PM significantly decreased maximal contractile responses
to norepinephrine (NE), but no change in EC50 was observed. In addition to
vascular study, the authors found no significant alteration in serum
cholesterol, LDL, triglyceride, HDL, ALT AST alkaline phosphatase, and lipid
peroxidation in OVX + PM rabbits. These data demonstrate that PM (100 mg/kg/d)
improved endothelial function through NO/dependent pathway and increased
response to E2 while sensitivity to NE was reduced. In addition, it had no
impact on lipid profile, liver enzymes, and ALP activities. PM is a potential
source of phytoestrogens for postmenopausal women to improve cardiovascular
function or reduce cardiovascular risks.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16862667 [PubMed / indexed for MEDLINE]
23: J Environ Biol. 2006 Jan;27(1):21/6.
Production of puerarin and isoflavones in cell suspension cultures of Pueraria
lobata (Willd.): effects of medium supplementation with casein hydrolysate and
coconut milk.
Li L, Zhang CR.
College of Life Science, South China Normal University, Guangzhou,
Guangdong/510631, China. liling@scnu.edu.cn
Callus induced from leaf explants of Pueraria lobata seedlings were suspended in
Gamborg B5 medium supplemented with 1 mg l(/1) 2,4/dichlorophenoxyacetic acid, 1
mg l(/1) naphthalene acetic acid, 0.5 mg l(/1) kinetin and 30 g l(/1) sucrose.
The effects of coconut milk and casein hydrolysate (CH) on cell growth and
yields of puerarin and isoflavones in cells suspension were studied. The
contents of total isoflavones and puerarin in suspension cultures were
determined by spectrophotometry and HPLC. Coconut milk (10%, filter sterilized)
decreased the growth of cell cultures and the accumulation of total isoflavones,
while 0.2% CH promoted the growth of cell cultures and the accumulation and
release of puerarin and total isoflavones. The total yield of puerarin and
isoflavones were 34% and 40.8% higher than in the control, respectively. The
optimum medium for cell cultures of leaves of P. lobata seedlings was B5 liquid
medium supplemented with 2% sucrose, 1.0 mg l(/1) 2,4/D, 1.0 mg l(/1) NAA, 0.5
mg l(/1) kinetin and 20 mg l(/1) CH. The procedure use is a potentially useful
for the production of isoflavones.
Publication Types:
Evaluation Studies
Research Support, Non/U.S. Gov't
PMID: 16850870 [PubMed / indexed for MEDLINE]
24: Plant Cell Rep. 2006 Dec;25(12):1387/91. Epub 2006 Jul 14.
Regulation of isoflavone production in hydroponically grown Pueraria montana
(kudzu) by cork pieces, XAD/4, and methyl jasmonate.
Kirakosyan A, Kaufman PB, Chang SC, Warber S, Bolling S, Vardapetyan H.
Department of Cardiac Surgery, University of Michigan, Ann Arbor, MI 48109/0686,
USA. pbk@umich.edu
A mini/hydroponic growing system was employed for seedlings of kudzu vine
(Pueraria montana) and contents of isoflavones (daidzein, genistein, daidzin,
genistin, and puerarin) from shoot and root parts of seedlings were analyzed
quantitatively. In addition, exogenous cork pieces, polymeric adsorbent, XAD/4,
and universal elicitor, methyl jasmonate (MeJA), were used to regulate the
production of these isoflavones. It was shown that cork pieces up/regulate the
production of daidzein and genistein up to seven/ and eight/fold greater than
the levels obtained for control roots. In contrast, levels of glucosyl
conjugates, daidzin and genistin, decrease up to five/ and eight/fold,
respectively. Cork treatment also induces the excretion of the root isoflavone
constituents into the growth medium. Minimal levels of isoflavones are absorbed
by the cork pieces. XAD/4 stimulates the production of glucosyl conjugates,
daidzin and genistin, in root parts about 1.5/fold greater than that obtained in
control roots. These are the highest amounts of daidzin and genistin that are
observed (5.101 and 6.759 mg g(/1) dry weight, respectively). In contrast to
these two adsorbents, MeJA increases the accumulation of isoflavones in shoot
rather than in root parts of seedlings, about three/ to four/fold over control
levels, with the exception of genistein. These studies reveal new observations
on the regulation of isoflavone production in hydroponically grown Pueraria
montana plants by two adsorbents (cork pieces and XAD/4) and MeJA elicitor.
Publication Types:
Research Support, N.I.H., Extramural
PMID: 16841218 [PubMed / indexed for MEDLINE]
25: J Ethnopharmacol. 2006 Nov 3;108(1):38/45. Epub 2006 Apr 28.
KIOM/79 inhibits LPS/induced iNOS gene expression by blocking NF/kappaB/Rel and
p38 kinase activation in murine macrophages.
Jeon YJ, Li MH, Lee KY, Kim JS, You HJ, Lee SK, Sohn HM, Choi SJ, Koh JW, Chang
IY.
College of Medicine, Chosun University, 375 Susukdong, Kwangju 501/709, Republic
of Korea.
We demonstrate that KIOM/79, combined extracts obtained from Magnolia
officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis,
inhibits LPS/induced expression of iNOS gene in RAW 264.7 cells. Treatment of
RAW 264.7 cells with KIOM/79 inhibited LPS/stimulated nitric oxide production in
a dose/related manner. Immunohisto/chemical staining of iNOS and RT/PCR analysis
showed that the decrease of NO was due to the inhibition of iNOS gene
expression. Immunostaining of p65, EMSA, and reporter gene assay showed that
KIOM/79 inhibited NF/kappa/Rel nuclear translocation, DNA binding, and
transcriptional activation, respectively. Western immunoblot analysis of p38
kinase showed KIOM/79 significantly inhibited the phosphoylation of p38 kinase
which is important in the regulation of iNOS gene expression. Collectively, this
series of experiments indicates that KIOM inhibits iNOS gene expression by
blocking NF/kappa/Rel and p38 kinase signaling. Due to the critical role that NO
release plays in mediating inflammatory responses, the inhibitory effects of
KIOM/79 on iNOS suggest that KIOM/79 may represent a useful anti/inflammatory
agent.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16806764 [PubMed / in process]
26: J Reprod Dev. 2006 Aug;52(4):537/42. Epub 2006 Jun 24.
Effect of Pueraria mirifica on the sexual skin coloration of aged menopausal
cynomolgus monkeys.
Trisomboon H, Malaivijitnond S, Cherdshewasart W, Watanabe G, Taya K.
Department of Physiology, Faculty of Medicine, Srinakharinwirot University,
Bangkok, Thailand.
To investigate the estrogenic effect of Pueraria mirifica (PM), a Thai herbal
plant that contains many phytoestrogens, sexual skin coloration was studied in
cynomolgus monkeys. Aged menopausal monkeys were divided into three groups. Each
group (n=3) was fed 10, 100, or 1,000 mg of PM daily. The treatment schedule was
divided into three periods, a 30/day pre/treatment period, 90/day treatment
period, and 60/day post/treatment period. The results show that the sexual skin
exhibited reddish coloration within 24 h after PM/treatment and remained this
way for the first half of the PM/feeding period. The changes in sexual skin
coloration were not dose/dependent. The present results indicate that PM had
estrogenic action by increasing reddish sexual skin coloration in aged
menopausal monkeys.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16799265 [PubMed / indexed for MEDLINE]
27: J Agric Food Chem. 2006 Jun 28;54(13):4907/12.
Both soybean and kudzu phytoestrogens modify favorably the blood lipoprotein
profile in ovariectomized and castrated hamsters.
Guan L, Yeung SY, Huang Y, Chen ZY.
Li Ka Shing Institute of Health Sciences, Food and Nutritional Sciences Program
of the Department of Biochemistry, and Department of Physiology, The Chinese
University of Hong Kong, Shatin, NT, Hong Kong, China.
The present study compared the hypolipidemic activity of kudzu phytoestrogens
with that of soybean phytoestrogen in estrogen/ and androgen/deficient hamsters.
In the first experiment, ovariectomized hamsters (n = 37) were randomly divided
into four groups (n = 9/10 each group). The first group was the control group,
whereas the second group had the time/releasing estradiol/17beta subcutaneous
(pellet) implants as a positive control. The third and fourth groups were orally
administered soybean or kudzu phytoestrogen extracts (30 mg/kg of body weight)
per day. In the second experiments, the first group of male hamsters (n = 9)
received a sham operation, whereas the other three groups of male hamsters (n =
9 each) were castrated. The castrated control group received orally distilled
water, whereas the second and third castrated groups were orally given 30 mg/kg
soybean or kudzu phytoestrogen extracts. The results for the first experiment
showed that the ovariectomized hamsters orally given soybean and kudzu
phytoestrogen extracts had significantly decreased serum total cholesterol (TC)
and non/high/density lipoprotein cholesterol (non/HDL/C) with HDL cholesterol
(HDL/C) being unaffected. The data from the second experiment demonstrated that
administration of soybean but not kudzu phytoestrogen extracts decreased
significantly serum TC. However, administration of kudzu phytoestrogens caused
redistribution of cholesterol among lipoproteins, leading to a significant
decrease in the ratio of non/HDL/C to HDL/C. It was concluded that both soybean
and kudzu phytoestrogens could modify favorably lipoprotein profiles in
ovariectomized and castrated hamsters.
PMID: 16787047 [PubMed / indexed for MEDLINE]
28: Zhongguo Zhong Yao Za Zhi. 2006 Apr;31(7):577/9.
[Protective effect of pueraria flavonoid on the cerebral ischemic reperfusion
injury in rats]
[Article in Chinese]
Wang PY, Wang HP, Li GW.
Department of Neurobiology, Anhui Medical University, Hefei 230032, China.
OBJECTIVE: To research the protective effect of pueraria flavonoid on the
cerebral ischemic reperfusion injury. METHOD: Using the middle cerebral artery
occlusion model (MCAO) in rats, we investigated the influence of pueraria
flavonoid on the brain water content, the infarct volume, the activities of SOD,
and the content of MDA. RESULT: Pueraria Flavonoid could obviously reduce the
brain water content and the infract volume in MCAO, increase the activities of
SOD, and decrease the content of MDA in the cerebral ischemia/ reinfusion model
of rats. CONCLUSION: Pueraria has the function of scavenging free radicals and
the protective effect on ischemic brain tissue.
Publication Types:
English Abstract
PMID: 16780163 [PubMed / in process]
29: J Cosmet Sci. 2006 Mar/Apr;57(2):201/2.
Botanicals from traditional chinese medicine (TCM) and their potential as
natural anti/irritants in cosmetic applications.
Vielhaber G, Meyer I, Joppe H, Franke H, Herrmann M.
Symrise GmbH & Co. KG, Muhlenfeldstr. 1, D/37603, Holzminden, Germany.
PMID: 16758567 [PubMed / indexed for MEDLINE]
30: Biol Pharm Bull. 2006 Jun;29(6):1202/6.
Estrogenic effect of main components kakkalide and tectoridin of puerariae flos
and their metabolites.
Shin JE, Bae EA, Lee YC, Ma JY, Kim DH.
College of Pharmacy, Kyung Hee University.
To understand the relationship between the metabolism and estrogenic activity of
kakkalide and tectoridin, main isoflavones in the flower of Pueraria
thunbergiana (family Leguminosae), these isoflavones and their metabolites by
human intestinal microflora as well as their estrogenic effects were
investigated. All human fecal specimens metabolized kakkalide and tectoridin.
All isolated kakkalide/hydrolyzing intestinal bacteria also hydrolyzed kakkalide
and tectoridin to irisolidone and tectorigenin, respectively. When the
estrogenic effects of kakkalide and tectoridin were compared with those of their
metabolites irisolidone and tectorigenin, the metabolites more potently
increased proliferation of MCF/7 cells than kakkalide and tectoridin. These
metabolites also potently induced estrogen/response c/fos and pS2 mRNA
expression. These results suggest that kakkalide and tectoridin may be
metabolized mainly to irisolidone and tectorigenin, respectively, by intestinal
microflora in the intestines, and which may be subsequently absorbed into the
blood where they can express their estrogenic effect.
PMID: 16755017 [PubMed / in process]
31: Ann Chim. 2006 Jan/Feb;96(1/2):117/24.
Separation and determination of isoflavonoids in several kudzu samples by
high/performance capillary electrophoresis (HPCE).
Fang C, Wan X, Tan H, Jiang C.
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and
Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, Anhui,
China.
Pueraria lobata is a rich source of isoflavonoids. The detection and
identification of isoflavonoid components from root, stem, leaf, callus and cell
samples, is very important for the best, safest and most efficacious use of
kudzu as a medicinal plant, and for the studies on quantitative analysis in the
secondary metabolism of isoflavonoids. In this paper, a simple, rapid and
precise high/performance capillary electrophoresis (HPCE) method with diode
array detection (DAD) has been developed for separation and determination of
isoflavonoids in several kudzu samples. The isoflavonoids could be well
separated within 15 min in a 40 cm length capillary at a separation voltage of
15kV in a 30 mmol L(/1) borax buffer (pH9.29), and this proposed method
demonstrated excellent reproducibility and accuracy with relative standard
deviations of less than 5% for isoflavonoid content (n = 5) of different kudzu
samples. The relationship between peak areas and isoflavone concentrations, in a
specified working range with linear response, was determined by first/order
polynomial regression over the range 0.05/0.5 mg mL(/1) for puerarin and 2.5/50
microg mL(/1) for 3'/methoxypuerarin, daidzin and daidzein, respectively, and
quantitative evaluation of those four main isoflavonoid components was
determined by ultraviolet absorption at lambda = 192 nm. The differences were
also illustrated by comparison of the determination of isoflavonoid components
from kudzu root, stem, leaf samples and plant tissue cultures in vitro.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16734027 [PubMed / indexed for MEDLINE]
32: J Ethnopharmacol. 2006 Oct 11;107(3):354/60. Epub 2006 Apr 6.
Using vaginal cytology to assess the estrogenic activity of phytoestrogen/rich
herb.
Malaivijitnond S, Chansri K, Kijkuokul P, Urasopon N, Cherdshewasart W.
Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn
University, Phyathai Road, Patumwan, Bangkok 10330, Thailand.
Suchinda.m@chula.ac.th
To assess the estrogenic activities of synthetic estrogen, synthetic
phytoestrogen, Pueraria lobata and three distinct cultivars of Pueraria
mirifica, a phytoestrogen/rich herb, a vaginal cytology assay in ovariectomized
rats were used. Rats were ovariectomized and treated with DW, estradiol valerate
(1 mg/kg BW), genistein (0.25/2.5 mg/kg BW), Pueraria lobata and Pueraria
mirifica (10/1,000 mg/kg BW) for 14 days. The vaginal cytology was checked daily
and the uteri were dissected and weighed at the end of treatment or
post/treatment periods. The treatments of DW, genistein and Pueraria lobata did
not influence the vaginal epithelium, but the injection of estradiol valerate
induced a vaginal cornification from day/3 of treatment to day/14 of
post/treatment period. The occurrence of vaginal cornification after treatment
and the recovery after the cessation was dependent on dosages and cultivars of
Pueraria mirifica. The increments of uterus weight in all rats agreed with the
cornification of vaginal epithelium. Although both uterotropic and vaginal
cytology assays can be used to assess the estrogenic activity of
phytoestrogen/rich herb, however, using vaginal cytology assay has two
advantages: (1) we do not need to kill the animals and (2) we can follow up the
recovery after the cessation of treatment.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
PMID: 16730147 [PubMed / indexed for MEDLINE]
33: Zhongguo Zhong Yao Za Zhi. 2006 Mar;31(6):504/6.
[Protective effect of puerarin on stress/induced gastric mucosal injury in rats]
[Article in Chinese]
Wang FW, Li J, Hu ZL, Xie YY.
Institute of Materia Medica, Shandong Academy of Medical Sciences, Jinan 250062,
China. wangfuwww@tom.com
OBJECTIVE: To study the protective effect of puerarin on stress/induced gastric
mucosal injury in rats. METHOD: The model of gastric ulcer was established by
restraint plus water/immersion stress in rats. Gastric motility was monitored by
the method of "Gas Balloon". Gastric mucosal blood flow was recorded by
laser/Doppler flowmetry. Colorimetric method was used to determine the content
of NO and ET in gastric mucosal tissue. Meantime the pathologic changes of
gastric mucosal was examined. RESULT: Puerarin could significantly attenuated
gastric mucosal damage induced by water/immersion stress, inhibited gastric
motility, specially decreased the index of gastric motility and percentage of
gastric contraction time and numbers of violent contraction. The gastric mucosal
blood flow and NO level in gastric mucosal were enhanced, while ET level was
reduced by puerarin. The degree of tissue damage in gastric mucosal was also
significantly attenuated after administration fo puerarin. CONCLUSION: Puerarin
exerts a significant protective effect on water/immersion stress/induced gastric
mucosal damage by relaxing the vessels, increasing NO level in gastric mucosal,
increasing regional gastric mucosal blood flow and inhibiting gastric motility.
Publication Types:
English Abstract
PMID: 16722386 [PubMed / indexed for MEDLINE]
34: Zhongguo Zhong Yao Za Zhi. 2006 Mar;31(6):490/3.
[Puerarin suppresses the proliferation of vscular smooth muscule cells and c/fos
and bcl/2 protein expression]
[Article in Chinese]
Xu YZ, Gao Y, Li PZ, Wang NF, Xu HY, Tong GX.
The First People's Hospital of Hangzhou, Hangzhou 310006, China.
qqyzxu@zj165.com
OBJECTIVE: To observe the role of puerarin on the proliferation of vascular
smooth muscle cells(VSMC) induced by thrombin (T) and the effect of puerarin on
the c/fos and bcl/2 protein expression. METHOD: Cell number and cell cycle
analysis using flow cytometry were adopted as two different indicators of
effects on proliferation of VSMC. Western blot was used to indicate the changes
of c/fos and bcl/2 protein after 24 h of treatment of T and puerarin. RESULT:
1.5 x 10(/5) / 1.5 x 10(/3) mol x L(/1) puerarin could significantly suppress
this stimulation of VSMC proliferation and DNA synthesis induced by T. Western
blot demonstrated that after 24 hour of treatment with T and puerarin, T could
significantly increase c/fos and bcl/2 protein and 1.5 x 10(/5) / 1.5 x 10(/3)
mol x L(/1) puerain could significantly suppress this increase. CONCLUSION:
puerarin can suppress the proliferation and DNA synthesis of VSMC promoted by T.
This inhibitory effects of puerarin are closely related with the suppression of
c/fos and bcl/2 protein.
Publication Types:
English Abstract
PMID: 16722382 [PubMed / indexed for MEDLINE]
35: J Chromatogr A. 2006 Jul 14;1121(1):114/9. Epub 2006 May 22.
High/performance thin/layer chromatographic fingerprints of isoflavonoids for
distinguishing between Radix Puerariae Lobate and Radix Puerariae Thomsonii.
Chen SB, Liu HP, Tian RT, Yang DJ, Chen SL, Xu HX, Chan AS, Xie PS.
State Key Laboratory of Chinese Medicine and Molecular Pharmacology, Department
of Applied Biology and Chemical Technology, The Hong Kong Polytechnic
University, Hung Hom, Hong Kong, China.
The roots of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth have been
officially recorded in all editions of Chinese Pharmacopoeia under the same
monograph 'Gegen' (Radix Puerariae, RP). However, in its 2005 edition, the two
species were separated into both individual monographs, namely 'Gegen' (Radix
Puerariae Lobatae, RPL) and 'Fenge' (Radix Puerariae Thomsonii, RPT),
respectively, due to their obvious content discrepancy of puerarin, the major
active constituent. In present paper, the fingerprint of high/performance
thin/layer chromatography (HPTLC) combining digital scanning profiling was
developed to identify and distinguish the both species in detail. The unique
properties of the HPTLC fingerprints were validated by analyzing ten batches of
Pueraria lobata and P. thomsonii samples, respectively. The common pattern of
the HPTLC images of the roots of Pueraria spp. and the respective different
ratios of the chemical distribution can directly discern the two species. The
corresponding digital scanning profiles provided an easy way for quantifiable
comparison among the samples. Obvious difference in ingredient content and HPTLC
patterns of the two species questioned their bio/equivalence and explained that
recording both species separately in the current edition of Chinese
Pharmacopoeia (2005 edition) is reasonable due to not only the content of major
constituent, puerarin, but also the peak/to/peak distribution in the fingerprint
and integration value of the total components. Furthermore, the HPTLC
fingerprint is also suitable for rapid and simple authentication and comparison
of the subtle difference among samples with identical plant resource but
different geographic locations.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16714027 [PubMed / indexed for MEDLINE]
36: Yao Xue Xue Bao. 2006 Feb;41(2):171/4.
[Protective effects of hydroxyethylpuerarin against brain astrocytes injury
induced by hydrogen peroxide]
[Article in Chinese]
Zhang B, Wei XB, Liu HQ, Wang LX, Sun R, Zhang XM.
Institute of Pharmacology, School of Medicine, Shandong University, Jinan
250012, China.
AIM: To study the protective effects of hydroxyethylpuerarin against the injury
of astrocytes induced by hydrogen peroxide (H2O2). METHODS: Experiments were
performed with cells from passage 4. Plasma membrane integrity was measured by
lactate dehydrogenase (LDH) release. The occurrence of apoptosis was measured by
flow cytometry. The glutamate uptake of astrocytes was studied with
[3H]/glutamate incorporation. Intracellular superoxide dismutase (SOD) activity
and malondialdehyde (MDA) level were assessed by automatic biochemistry
analyzer. RESULTS: Compared with H2O2 injured group, the occurrence of
apoptosis, levels of LDH release and intracellular MDA of astrocytes reduced in
hydroxyethylpuerarin pre/treated groups, but the glutamate uptake and
intracellular SOD activity of astrocytes increased. CONCLUSION:
Hydroxyethylpuerarin could reduce the occurrence of apoptosis and improve
neurotrophic function of astrocytes, which may be related with its antioxidant
effects during oxidative stress.
Publication Types:
English Abstract
PMID: 16671550 [PubMed / indexed for MEDLINE]
37: Endocrine. 2006 Feb;29(1):129/34.
The estrogenic effect of Pueraria mirifica on gonadotrophin levels in aged
monkeys.
Trisomboon H, Malaivijitnond S, Watanabe G, Cherdshewasart W, Taya K.
Department of Physiology, Faculty of Medicine, Srinakharinwirot University,
Bangkok 10110, Thailand.
We investigated the effect of Pueraria mirifica (PM) on gonadotrophin and
estradiol levels in aged animals; nine menopausal cynomolgus monkeys were
divided into three groups. Each group (n = 3) was fed with 10, 100, and 1000
mg/d of PM for 90 d. PM/10 induced the decrease of follicle stimulating hormone
(FSH) levels on d 15/90 in one out of three monkeys. PM/100 and PM/1000
decreased FSH levels of all monkeys throughout the treatment period. After the
treatment period, FSH levels continued to decrease for 5 and 10/20 d in PM/100
and PM/1000, respectively, and the levels rebounded in all groups thereafter.
PM/10 decreased luteinizing hormone (LH) levels throughout the treatment period
in one out of three monkeys and returned to the pretreatment levels immediately
after stopping treatment. PM/100 and PM/1000 prominently decreased LH levels
between d 10 and 90 during treatment and persisted until d 15/25 and d 20/30 for
PM/100 and PM/1000, respectively, during the post/treatment period. Serum LH
levels rebounded after returning to pre/treatment levels in a dose/dependent
manner. Estradiol levels tended to decrease during the treatment period in all
groups. The daily feeding of PM suppressed gonadotrophin levels in aged
menopausal monkeys based on dose. Moreover, they can be recovered, and there is
a direct correlation between dosage and recovery time. PM may be effective as an
alternative medicine in menopausal women because the effects are not permanent.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16622301 [PubMed / indexed for MEDLINE]
38: J Chromatogr Sci. 2006 Feb;44(2):57/63.
Identification of isoflavonoids in several kudzu samples by high/performance
liquid chromatography coupled with electrospray ionization tandem mass
spectrometry.
Fang C, Wan X, Tan H, Jiang C.
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and
Ministry of Agriculture, P.R. China.
Pueraria lobata is a rich source of isoflavonoids. The detection and
identification of isoflavonoid components from Pueraria radix (RP), callus and
cell cultures, is very important for the safest and most effective use of kudzu
as a medicinal plant, and for the studies on quantitative analysis and secondary
metabolism of isoflavonoids in vitro cultures. Liquid chromatography is coupled
with negative and positive electrospray ionization (ESI) tandem mass
spectrometry (MS/MS), and photodiode array detection is used to characterize and
detect isoflavonoids in root, callus, and cell samples of P. lobata.
Characteristic product ions of aglycones, O/glucosides, and C/glucosides were
obtained from the full/scan ESI/MS chromatography of the major peaks and the
MS/MS spectra of the protonated ions. Five major components of puerarin,
daidzin/6"/O/acetylester, genistin/6"/O/malonylester, biochanin
A/7/O/glucoside/6"/O/malonylester, and daidzein are detected and identified from
the methanolic extract of P. lobata callus cultures. The major isoflavonoid
components of P. lobata cell suspension cultures are identified as puerarin,
daidzin, daidzin/6"/O/acetylester, genistin/6"/O/malonylester, biochanin
A/7/O/glucoside/6"/O/malonylester, genistein/8/C/glucoside/6"/O/malonylester,
and daidzein, on the basis of ESI/MS and MS/MS spectra analysis. Likewise,
puerarin, daidzin, genistein/6"/O/malonylester, 3'/methoxypuerarin, and daidzein
are detected and identified from RP. Of those isoflavonoid components detected,
daidzin/6"/O/acetylester is a new isoflavonoid glucoside and is for the first
time detected from P. lobata cultures in vitro.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16620495 [PubMed / indexed for MEDLINE]
39: J Nutr. 2006 Apr;136(4):899/905.
Differential effects of isoflavones, from Astragalus membranaceus and Pueraria
thomsonii, on the activation of PPARalpha, PPARgamma, and adipocyte
differentiation in vitro.
Shen P, Liu MH, Ng TY, Chan YH, Yong EL.
Department of Obstetrics and Gynecology, Yong Loo Lin School of Medicine,
National University of Singapore, 119074, Singapore.
Compounds that target the peroxisome proliferator/activated receptors PPARalpha
and PPARgamma are used to correct dyslipidemia and to restore glycemic balance,
respectively. Because the majority of diabetic patients suffer from atherogenic
lipid abnormalities, in addition to insulin resistance, ligands are required
that can activate both PPARalpha and PPARgamma. In this study, we used chimeric
PPARalpha/gamma reporter/gene bioassays to screen herbal extracts with purported
antidiabetic properties. Extracts of Astragalus membranaceus and Pueraria
thomsonii significantly activated PPARalpha and PPARgamma. Bioassay/guided
fractionation resulted in the isolation of the isoflavones, formononetin, and
calycosin from Astragalus membranaceus, and daidzein from Pueraria thomsonii as
the PPAR/activating compounds. We investigated the effects of these and 2 common
isoflavones, genistein and biochanin A, using chimeric and full/length PPAR
constructs in vitro. Biochanin A and formononectin were potent activators of
both PPAR receptors (EC50 = 1/4 micromol/L) with PPARalpha/PPARgamma activity
ratios of 1:3 in the chimeric and almost 1:1 in the full/length assay,
comparable to those observed for synthetic dual PPAR/activating compounds under
pharmaceutical development. There was a subtle hierarchy of PPARalpha/gamma
activities, indicating that biochanin A, formononetin, and genistein were more
potent than calycosin and daidzein in chimeric as well as full/length receptor
assays. At low doses, only biochanin A and formononetin, but not genistein,
calycosin, or daidzein, activated PPARgamma/driven reporter/gene activity and
induced differentiation of 3T3/L1 preadipocytes. Our data suggest the potential
value of isoflavones, especially biochanin A and their parent botanicals, as
antidiabetic agents and for use in regulating lipid metabolism.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
PMID: 16549448 [PubMed / indexed for MEDLINE]
40: Langmuir. 2006 Mar 14;22(6):2827/31.
Easy replication of Pueraria Lobata toward hierarchically ordered porous
gamma/Al2O3.
Li C, He J.
Functional Nanomaterials Laboratory, Technical Institute of Physics and
Chemistry, Chinese Academy of Sciences (CAS), Chaoyangqu Datunlu Jia 3, Beijing
100101, China.
Hierarchically ordered porous alumina was prepared via a facile
immersion/fuming/calcination process using Pueraria lobata as template. The
as/prepared alumina inherited nearly all morphological features of the template,
as shown by SEM observations. It also contains abundant mesopores based on
nitrogen adsorption/desorption measurements. The crystalline phase of the
as/prepared alumina was ascertained to be gamma/alumina by analyzing its XRD
pattern. Pt nanoparticles were in situ synthesized in the gamma/alumina matrix
and annealed at different temperatures in N(2) atmosphere. TEM observations
showed that Pt nanoparticles supported by the as/prepared alumina have
significantly high thermal stability.
PMID: 16519490 [PubMed]
41: Sheng Wu Gong Cheng Xue Bao. 2005 Nov;21(6):1003/8.
[Effects of sucrose and light on the growth and production of secondary
metabolites in Pueraria phaseoloides hairy roots]
[Article in Chinese]
He HJ, Liang P, Shi HP.
Guangdong Key Lab of Biotechnology for Plant Development, College of Life
Science, South China Normal University, Guangzhou 510631, China.
Effects of sucrose concentrations and light on the growth and production of
total isoflavones and puerarin in Pueraria phaseoloides hairy roots cultured
onto solid MS media supplemented with 1%, 3%, 5%, 7% and 9% sucrose,
respectively, were investigated. The results showed that among the sucrose
concentrations tested, 3% sucrose in the medium enhanced the growth and
stimulated accumulation of total isoflavones and puerarin in P. phaseoloides
hairy roots, After cultured for 20 days, the biomass of hairy roots reached 0.48
g (DW dry weight)/flask and its contents of total isoflavones and puerarin were
25.44 mg/g (DW) and 11.64 mg/g (DW), respectively. In comparison with 3%
sucrose, the dry weight proliferation of hairy roots cultured with 5% sucrose
was increased by 7.0%, while cultured with 1%, 7% and 9% sucrose, the dry weight
proliferation of hairy roots was decreased by 62.4%, 42.8% and 65.3%, their
total isoflavones content was decreased by 57.4%, 13% and 33.4% and their
puerarin content was decreased by 47.9%, 15.8% and 35.1%; but their content of
total soluble sugars was increased 0.52, 1.45 and 1.54 times, respectively.
Compared with hairy roots in blue light and white light, the biomass of hairy
roots cultured in the dark for 30 days was 0.83 g (DW)/flask and was increased
by 37.1% and 23.3%, respectively. The content of total isoflavones in hairy
roots cultured in white light was as much as 1.15 times and 1.19 times that in
blue light and in the dark, respectively. It was also observed that hairy roots
cultured in blue light and white light partly became light green and that blue
light could inhibit accumulation of puerarin in hairy roots and the puerarin
content in hairy root cultured in white light and in the dark were 1.61 times
and 1.52 times that in blue light, respectively.
Publication Types:
English Abstract
PMID: 16468362 [PubMed / in process]
42: J Ethnopharmacol. 2006 Apr 6;104(3):345/50. Epub 2006 Feb 7.
Puerariae radix promotes differentiation and mineralization in human
osteoblast/like SaOS/2 cells.
Huh JE, Yang HR, Park DS, Choi DY, Baek YH, Cho EM, Cho YJ, Kang/Il K, Kim DY,
Lee JD.
Oriental Medicine Research Center for Bone & Joint Disease, Kyung Hee
University, 1 Hoegidong, Dongdaemungu, Seoul 130/701, Republic of Korea.
Puerariae radix (PR) is a traditional medicine herb used for enhancing body
resistance against various diseases. The aim of this study was to identify
whether Puerariae radix extract induces osteogenic activity in human
osteoblast/like SaOS/2 cells. Puerariae radix had no effect on the viability of
osteoblastic cells, and dose/dependently increased alkaline phosphatase (ALP)
activity. Puerariae radix markedly increased mRNA expression for vascular
endothelial growth factor (VEGF), osteocalcin (OCN), osteopontin (OPN), and type
I collagen (Col I) in SaOS/2 cells. Extracellular accumulation of proteins such
as VEGF and Col I was increased in a dose/dependent manner. Also, Puerariae
radix significantly induced mineralization in the culture of SaOS/2 cells. In
conclusion, this study showed that Puerariae radix had no effect on viability,
but enhanced ALP activity, VEGF, bone matrix proteins such as OCN, OPN, and Col
I, and mineralization in SaOS/2 cells. These results propose that Puerariae
radix can play an important role in osteoblastic bone formation, and may
possibly lead to the development of bone/forming drugs.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16455216 [PubMed / indexed for MEDLINE]
43: Southeast Asian J Trop Med Public Health. 2005;36 Suppl 4:238/41.
Mutagenicity of Pueraria mirifica Airy Shaw & Suvatabandhu and antimutagenicity
of Thunbergia laurifolia Linn.
Saenphet K, Kantaoop P, Saenphet S, Aritajat S.
Biology Department, Faculty of Science, Chiang Mai University, Chiang Mai 50200,
Thailand. K_Saenphet@yahoo.com
Thunbergia laurifolia Linn has been reputed to have antitoxic effects for all
toxic substances. In this present study, we evaluated its effect against the
mutagenicity induced by aqueous extracts from Pueraria mirifica Airy Shaw &
Suvatabundhu in male rats. The formation of micronuclei in polychromatic
erythrocytes was induced by oral administration of an aqueous extract of P.
mirifica at the doses of 400, 600, and 800 mg/kg to the rats for 30 days. The
results were that the extracts of P. mirifica at doses of 600 and 800 mg/kg
acted as a mutagenic agent by inducing higher frequencies of micronuclei as
compared to the controls. For the antimutagenic test, P. mirifica extract at a
dose of 600 mg/kg (minimal effective dose) was mixed with fresh and dried
extracts of T. laurifolia in proportions of 7:3 and 1:1, respectively. The
results of 4/week/treatment indicated that aqueous extracts of T. laurifolia,
prepared by both fresh and dry methods, could significantly inhibit the
induction of micronuclei as induced by P. mirifica. It could be concluded from
the results that, under certain circumstances, T. laurifolia exhibits a
significant antimutagenic activity. The use of P. mirifica and T. laurifolia as
fusion herbal medicines is suggested.
PMID: 16438216 [PubMed / indexed for MEDLINE]
44: Southeast Asian J Trop Med Public Health. 2005;36 Suppl 4:167/75.
Three indigenous Thai medicinal plants for control of Aedes aegypti and Culex
quinquefasciatus.
Lapcharoen P, Apiwathnasorn C, Komalamisra N, Dekumyoy P, Palakul K, Rongsriyam
Y.
Insecticide Research Unit, Department of Medical Entomology, Faculty of Tropical
Medicine, Mahidol University, Bangkok, Thailand. neparichart@yahoo.com
The potential larvicidal activity and insect growth regulator (IGR) properties
of three selected indigenous medicinal Thai plants were tested against two
species of mosquito with special reference to the late 3rd and early 4th instar
larvae (L3 and L4, respectively). In case of larvicidal activity, Thevetia
peruviana was the most potent, followed by Pueraria mirifica, and Butea superba
was the least effective. In all cases, the late 3rd instar was more susceptible
than the early 4th instar larvae, and the 48/hours exposure yielded more potent
larvicidal activity than 24/hours exposure. However, at sublethal dosages, both
P. mirifica and B. superba showed some dispersed effects interfering with
ecdysis. A variety of toxic effects were observed and recorded in eight
categories according to the stage of metamorphosis when death occurred. P.
mirifica rendered the main deleterious effects in the pupa/adult period in both
instar of Ae. aegypti and Cx. quinquefasciatus, whereas B. superba showed
highest effect in black/pupa period of the late 3rd instar larval stage. The
results were reversed for the early 4th instar larvae of both species of
mosquito as the main effect appeared in the pupa/adult category. The overall
results indicated that T. peruviana did not show any IGR properties; whereas, P.
mirifica and B. superba seemed to exhibit the juvenile hormone type activity
which resulted in abnormal death at various stages of development. B. superba
was more promising than P. mirifica, and Ae. aegypti was about 2 times more
susceptible than Cx. quinquefasciatus. In addition, L3 was always more
susceptible than L4 with both mosquito species.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16438204 [PubMed / indexed for MEDLINE]
45: Biosci Biotechnol Biochem. 2006 Jan;70(1):107/11.
Daidzein activates choline acetyltransferase from MC/IXC cells and improves
drug/induced amnesia.
Heo HJ, Suh YM, Kim MJ, Choi SJ, Mun NS, Kim HK, Kim E, Kim CJ, Cho HY, Kim YJ,
Shin DH.
Jeonnam Innovation Agency, Jeonnam, Korea.
The choline acetyltransferase (ChAT) activator, which enhances cholinergic
transmission via an augmentation of the enzymatic production of acetylcholine
(ACh), is an important factor in the treatment of Alzheimer's disease (AD).
Methanolic extracts from Pueraria thunbergiana exhibited an activation effect
(46%) on ChAT in vitro. Via the sequential isolation of Pueraria thunbergiana,
the active component was ultimately identified as daidzein
(4',7/dihydroxy/isoflavone). In order to investigate the effects of daidzein
from Pueraria thunbergiana on scopolamine/induced impairments of learning and
memory, we conducted a series of in vivo tests. Administration of daidzein (4.5
mg/kg body weight) to mice was shown significantly to reverse
scopolamine/induced amnesia, according to the results of a Y/maze test.
Injections of scopolamine into mice resulted in impaired performance on Y/maze
tests (a 37% decreases in alternation behavior). By way of contrast, mice
treated with daidzein prior to the scopolamine injections were noticeably
protected from this performance impairment (an approximately 12%/21% decrease in
alternation behavior). These results indicate that daidzein might play a role in
acetylcholine biosynthesis as a ChAT activator, and that it also ameliorates
scopolamine/induced amnesia.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16428827 [PubMed / indexed for MEDLINE]
46: J Chromatogr A. 2006 Feb 24;1107(1/2):59/69. Epub 2006 Jan 18.
Determination of isoflavones in dietary supplements containing soy, Red Clover
and kudzu: extraction followed by basic or acid hydrolysis.
Delmonte P, Perry J, Rader JI.
Office of Nutritional Products, Labeling and Dietary Supplements, Center for
Food Safety and Applied Nutrition, Food and Drug Administration, College Park,
MD 20740, USA. pierluigi.delmote@cfsan.fda.gov
Isoflavones are phytochemicals found in many plants. Because of their structural
similarity to beta/estradiol, health benefits of isoflavones have been evaluated
in age/related and hormone/dependent diseases. Dietary supplement preparations
contain extracts from soy, Red Clover and kudzu. Soy products contain primarily
genistein, daidzein, and glycitein, while Red Clover products contain primarily
formononetin and biochanin A. Kudzu extracts contain puerarin and daidzein among
other components. Previous methods of analysis focused on the determination of
isoflavones from a single botanical source, while dietary supplements are often
a blend of extracts from different plants. We developed a method for the
analysis of isoflavones in dietary supplements regardless of their botanical
composition, using HPLC/PDA because of its applicability to routine analysis.
Isoflavones are found as free compounds, glucoside derivatives,
6''/O/malonyl/beta/d/glucoside and 6''/O/acetyl/beta/d/glucoside derivatives. In
this study, the samples were extracted at room temperature with 50:50 (v/v)
MeCN/water, and then analyzed before and after hydrolyzing the isoflavones by
acid or basic digestion. 2'/Methoxy/flavone and 6/methoxy/flavone were used as
internal standards and were added together to every sample. Daidzein, glycitein,
genistein, puerarin, calycosin, pratensein, pseudobaptigenin, formononetin,
biochanin A and prunetin were among the isoflavones determined.
PMID: 16413562 [PubMed / indexed for MEDLINE]
47: Guang Pu Xue Yu Guang Pu Fen Xi. 2005 Sep;25(9):1507/9.
[Determination of copper, zinc, iron, calcium and magnesium in pueraria lobata
ohwi by FAAS]
[Article in Chinese]
Han P, Liu LE, Liu J, Zhang HQ, Wu YM, Wu YJ, Yu F, Wu DS.
College of Public Health, Zhengzhou University, Zhengzhou 450052, USA.
The samples of pueraria lobata ohwi were digested by HNO3 + HClO4. Copper, zinc,
iron, calcium and magnesium in pueraria lobata ohwi from Da Bieshan were
successfully determined by flame atomic absorption spectrophotometry (FAAS). The
results showed that there are rich grand elements, such as Ca and Mg, and
profitable elements, such as trace elements Cu, Zn and Fe, in pueraria lobata
ohwi. The content sequence of metal elements is as follows: Copper: leaves >
flowers > rattans > roots; Zinc: leaves > flowers > rattans > roots; Iron: roots
> rattans > leaves > flowers; Calcium: leaves > roots > flowers > rattans;
Magnesium: flowers > roots > leaves > rattans. The results provided useful data
for discussing the relationship between the metal elements in pueraria lobata
and hypertension diseases. ohwi and the cure for cerebrovascular diseases.
Publication Types:
English Abstract
PMID: 16379303 [PubMed / in process]
48: Drug Alcohol Rev. 2005 Nov;24(6):525/36.
Comment in:
Drug Alcohol Rev. 2005 Nov;24(6):473.
Natural medicines for alcoholism treatment: a review.
Xu BJ, Zheng YN, Sung CK.
Department of Food Science and Technology, College of Agriculture and
Biotechnology, Chungnam National University, Daejon, South Korea.
Alcoholism is a serious problem throughout the world. The development of
alcoholism remedies have medical, social and economical significance. In view of
the pitfalls of psychological dependence and adverse behavioural effects of
synthetic drugs, the development of low toxicity and high efficiency medicines
derived from natural products exhibits expansive market prospects. Based on
these considerations, we summarize briefly folk application of traditional
hangover remedies and clinical application of herbal complex and patent
medicines for alcoholism treatment. We have reviewed the effects of natural
medicines on intake, absorption and metabolism of alcohol, as well as the
protective effects on alcohol/induced acute and chronic tissue injury.
Publication Types:
Review
PMID: 16361209 [PubMed / indexed for MEDLINE]
49: Se Pu. 2005 Sep;23(5):477/81.
[Separation and identification of isoflavonoids in Pueraria lobata extracts and
its preparations by reversed/phase capillary liquid chromatography coupled with
electrospray ionization quadrupole time of flight mass spectrometry]
[Article in Chinese]
Tian H, Wang H, Guan Y.
Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, Dalian
116023, China.
Isoflavonoids in Pueraria lobata extract and its preparations were separated and
identified by reversed/phase capillary liquid chromatography (RP/CapLC) coupled
with photodiode array (PDA) detector and negative electrospray ionization
quadrupole time of flight (Q/TOF) mass spectrometry. The separation was
performed on a 150 mm x 0. 32 mm i. d. , 5 microm C18 capillary column, using
mobile phase of 0.1% aqueous trifluoroacetic acid solution and acetonitrile
containing 0.1% trifluoroacetic acid under gradient elution. The product ion
spectra of the deprotonated ions allowed for the identification of puerarin,
daidzin and daidzein in the sample. Puerarin was found to be the most abundant
component in the extract (about 13%, mass fraction) and its preparations (19.28
/ 24.34 mg per tablet). The structures of trace amount of unknown isoflavonoids
were deduced based on the spectra of known compounds. They were proposed to be
3'/methoxypuerarin and 3'/methoxydaidzin.
Publication Types:
English Abstract
PMID: 16350789 [PubMed / in process]
50: J Toxicol Environ Health A. 2005 Dec 10;68(23/24):2219/26.
Altered oral absorption of alcohol by combined aqueous extracts of four herbal
plants in rats.
Shin BS, Jun H, Lee DE, Lee KR, Park ES, Yoo SD.
College of Pharmacy, Sungkyunkwan University, Changan/gu, Suwon, Kyonggi/do,
Korea.
This study examined the effect of combined aqueous extracts (BHR) of Ginko
biloba, Mentha arvensis var. piperascens, Citrus unshiu, and Pueraria lobata
var. chinensis on oral absorption of alcohol in rats. The rats were pretreated
with BHR, placebo solution identical to BHR without the herbal extract, and
isotonic saline. Alcohol was administered orally at 1/ and 3/g/kg doses and the
absorption profiles were compared. After oral administration of 1/g/kg doses,
mean area under the curve (AUC) and C(max) values were significantly reduced in
BHR/treated rats (16.1 +// 10.0 and 0.3 +// 0.1 mg/ml, respectively) as compared
with saline/treated (37.9 +// 14.4 and 0.7 +// 0.7 mg/ml, respectively) and
placebo solution/treated (63.0 +// 46.4 and 0.7 +// 0.4 mg/ml, respectively)
rats. Similarly, after administration of 3/g/kg doses, mean AUC and C(max)
values in BHR/treated rats (188.1 +// 119.7 mg(.)min/ml and 1.0 +// 0.4 mg/ml)
were significantly reduced over those in saline/treated rats (571.4 +// 512.4
mg(.)min/ml and 1.8 +// 0.9 mg/ml, respectively). The relative oral
bioavailability of alcohol calculated as the ratio of AUC(BHR)/AUC(Saline) was
42.5% and 32.9% at 1/ and 3/g/kg doses, respectively. The reduced serum alcohol
levels as well as the reduced AUC and C(max) after pretreatment with BHR appear
to be a result of a reduced systemic absorption not due to an increased
metabolic clearance.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16326435 [PubMed / indexed for MEDLINE]
51: Zhonghua Yi Xue Za Zhi. 2005 Nov 2;85(41):2930/3.
[Effect of pueraria crude extreact and puerarin on ethanol/induced expression of
heat shock protein 70 in embryonic mouse hippocampal cultures]
[Article in Chinese]
Han P, Wu DS, Li WJ, Yu ZL, Wang Q.
Huaxi Public Health School, Sichuan University, Chengdu 610041, China.
OBJECTIVE: To study if the Pueraria crude extreact (CP) and standard preparation
of pure puerarin (SP) possess the same neuroprotective effects on the expression
of heat shock protein (HSP) 70 in the embryonic mouse hippocampal cells.
METHODS: The hippocampus of 18/days/old mouse embryo was taken out and
suspension of single cells was cultured. Ethanol was added to cause HSP70 mRNA
expression. Solvent, ethanol of different concentrations (50, 200, and 300
mmol/L), SP + ethanol, and SP + ethanol were added respectively. Western
blotting was used to detect the expression of the expression of HSP70 mRNA.
RESULTS: Ethanol of different concentrations increased the expression of HSP70
mRNA and the protein in comparison with the solvent control group. SP and CP
inhibited the expression of HSP70 mRNA and protein. CONCLUSION: With identical
effect of anti/oxidative stress, both SP and CP inhibit the increase of
expression of HSP70 mRNA and protein, thus demonstrating I vitro anti/oxidative
neuroprotection.
Publication Types:
English Abstract
PMID: 16324368 [PubMed / in process]
52: Alcohol Clin Exp Res. 2005 Oct;29(10):1913; author reply 1914.
Comment on:
Alcohol Clin Exp Res. 2005 May;29(5):756/62.
Is herbal "root" effective for reducing alcohol drinking?
Hayashino Y.
Publication Types:
Comment
Letter
PMID: 16269923 [PubMed / indexed for MEDLINE]
53: Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Sep;19(3):244/7.
[Potential antiviral drug pueraria crude extract and puerarin protect against
ethanol/induced cytotoxicity in embryonic mouse hippocampal cultures]
[Article in Chinese]
Han P, Li J, Li WJ, Yu ZL, Wang Q, Wu DS.
Huaxi School of Public Health, Sichuan University, Chengdu 610041, China.
OBJECTIVE: To examine whether Chinese medical herb pueraria crude extract (CP)
and standard of pure puerarin (SP) possess the same neuroprotective effects
during concomitant ethanol (EtOH) treatment. METHODS: Hippocampus cultures were
prepared from mice at gestational age of 18 day. Cell viability was measured by
MTT assay. RT/PCR was employed to determine mRNA expression of superoxide
dismutase (SOD). RESULTS: As measured by MTT assay, supplementation with 15 mg/L
CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50,
200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In
addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at
mRNA level. CONCLUSION: This study suggests that CP and SP could decrease
oxidative stress induced by ethanol treatment by the decreased expression of SOD
at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and
SP against developmental ethanol exposure in vitro.
Publication Types:
English Abstract
Research Support, Non/U.S. Gov't
PMID: 16261207 [PubMed / in process]
54: Mol Nutr Food Res. 2005 Dec;49(12):1154/9.
Suppression of arachidonic acid metabolism and nitric oxide formation by kudzu
isoflavones in murine macrophages.
Jun M, Hong J, Jeong WS, Ho CT.
Department of Food Science, Rutgers University, 65 Dudley Road, New Brunswick,
NJ 08901/8520, USA.
Inhibitory effect of kudzu isoflavones on arachidonic acid metabolism and nitric
oxide (NO) production in lipopolysaccharide activated RAW 264.7 macrophages were
investigated. Isoflavone aglycones, such as daidzein, genistein, biochanin A,
and formononetin significantly suppressed arachidonic acid release (50 microM).
Biochanin A, which displayed the most active inhibition on arachidonic acid
release in HT/29 human colon cancer cells, exhibited its most potent suppression
in RAW 264.7 cell (by 86%) without showing cytotoxicity. However, isoflavone
glucosides, puerarin and daidzin, showed lower inhibitory activities on the
release of arachidonic acid and its metabolites. In NO formation, biochanin A
showed marked inhibition, by 62% (50 microM), followed by genistein, daidzein,
formononetin, and daidzin, 56, 39, 33, and 8%, respectively. 5,7/Dihydroxyl
group in the A/ring of isoflavones could be a key functional group responsible
for the strong inhibitory activity of biochanin A and genistein on NO
production. These activities may contribute to the antiinflammatory and
anticarcinogenic properties of kudzu isoflavones.
PMID: 16254887 [PubMed / indexed for MEDLINE]
55: Metabolism. 2005 Nov;54(11):1536/41.
Puerariae radix prevents bone loss in castrated male mice.
Wang X, Wu J, Chiba H, Yamada K, Ishimi Y.
Division of Applied Food Research, National Institute of Health and Nutrition,
Tokyo 162/8636, Japan.
Puerariae radix (PR) is one of the earliest and most important crude herbs used
in Chinese medicine for various medicinal purposes. PR contains a high amount of
isoflavonoids, such as daidzein and genistein, which are known to prevent bone
loss induced by estrogen deficiency. We have demonstrated that PR not only
completely prevents bone loss but also significantly increases the bone mass at
high doses in ovariectomized mice without exhibiting estrogenic action in the
uterus. In this study, we examined whether PR exhibits effects on bone loss in
androgen/deficient male mice similar to estrogen/deficient female mice. Male
mice were orchidectomized (ORX) and fed a diet containing low, middle, and high
doses (5%, 10%, and 20% of diet, respectively) of PR or normal diet with
subcutaneous administration of 17beta/estradiol (E(2), 0.03 microg/d; Sigma, St
Louis, Mo), for 4 weeks. In ORX mice, the seminal vesicle weight decreased
markedly, and it was not affected by the administration of any doses of PR and
E(2). The bone mineral density (BMD) of the whole femur was significantly
decreased by ORX, and the decrease in BMD was completely prevented by intake of
the diet with the low dose of PR. Intake of the diet with the middle dose of PR
further normalized BMD in ORX mice. Furthermore, the high dose of PR
administration (PR20) significantly increased BMD in ORX mice, and the potency
was similar to that of E(2). Morphometric analysis of the femoral metaphysis
showed that intake of the diet with the low dose of PR completely prevented the
decrease in bone volume/tissue volume and trabecular number and restored the
increase in trabecular separation in ORX mice. In addition, intake of the diet
with the high dose of PR further increased bone volume/tissue volume and
trabecular number and decreased trabecular separation in ORX mice. These results
propose the possibility that estrogenic Chinese herbs such as PR can be one of
the candidates for the treatment or prevention of osteoporosis in elderly men
with hypogonadism.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16253645 [PubMed / indexed for MEDLINE]
56: Anal Bioanal Chem. 2005 Nov;383(5):787/96. Epub 2005 Nov 5.
High/performance liquid chromatography/tandem mass spectrometry for
identification of isoflavones and description of the biotransformation of kudzu
root.
Zhang Y, Xu Q, Zhang X, Chen J, Liang X, Kettrup A.
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian,
116023, PR China.
High/performance liquid chromatography/tandem mass spectrometry has been used to
identify isoflavone aglycones and glycosides in kudzu root. Fourteen isoflavones
were detected. Among these, six were identified by comparison with authentic
standards. Tentative identifications of the other isoflavones are based on UV
spectra, mass spectra of protonated and deprotonated molecules, and MS/MS data.
Several are reported for the first time in kudzu root. The bioactivity and
bioavailability of isoflavone aglycones are usually greater than those of their
glycosides. To improve the bioavailability of kudzu root isoflavones, crude
beta/glycosidases prepared from microbes were used to hydrolyze the isoflavone
glycosides. Several MS modes are combined not only to identify the isoflavones
in kudzu root, but also to describe the biotransformation of kudzu root
isoflavone glycosides. It is also proved that crude beta/glycosidases have high
selectivity toward the O/glycosides of isoflavones.
Publication Types:
Evaluation Studies
Research Support, Non/U.S. Gov't
PMID: 16237543 [PubMed / indexed for MEDLINE]
57: Harv Womens Health Watch. 2005 Sep;13(1):6/7.
Herbal treatment helps curb urge to drink.
[No authors listed]
Publication Types:
News
PMID: 16229064 [PubMed / indexed for MEDLINE]
58: J Sep Sci. 2005 Sep;28(14):1785/95.
Determination of 12 pueraria components by high/performance liquid
chromatography/mass spectrometry.
Lin CC, Wu CI, Sheu SJ.
Department of Chemistry, National Taiwan Normal University, Taipei, Taiwan, P.R.
China. chingchekimo@yahoo.com.tw
Puerariae radix, a commonly used Chinese herb drug derived from the dried root
of legume plant, contains a series of isoflavones as its chief pharmacologically
active constituents. Using 12 pueraria components as markers, an LC/UV/MS method
requiring less than 60 min, was developed for estimating the quality of pueraria
samples within 60 min. Extracts were analyzed using a Cosmosil 5C18/MS column,
by gradient elution with an aqueous solution of acetic acid and methanol/ACN at
a flow/rate of 1.0 mL/min. Peaks were detected at 254 nm and each peak was
identified by LC/MS. The reproducibilities (RSD) of this proposed method, on the
basis of peak/area ratios from six replicate injections, were 0.93/1.42%
(intraday) and 1.05/1.63% (interday) at a detection limit of 0.12/0.78
microg/mL. Most of the markers used in this study can be classified,
respectively, into three major categories, namely, isoflavones, O/glycosidic
isoflavones, and C/glycosidic isoflavones. The structures of the compounds were
determined from LC/MS fragmentation data and data from the literature.
Publication Types:
Research Support, Non/U.S. G
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