Pueraria thomsonii has long been used in traditional Chinese medicine. Isoflavonoids are the principle pharmacologically active components, which are primarily observed as glycosyl-conjugates and accumulate in P. thomsonii roots. However, the molecular mechanisms underlying the glycosylation processes in (iso)flavonoid biosynthesis have not been thoroughly elucidated. In the current study, an O-glucosyltransferase (PtUGT8) was identified in the medicinal plant P. thomsonii from RNA-seq database. Biochemical assays of the recombinant PtUGT8 showed that it was able to glycosylate chalcone (isoliquiritigenin) at the 4-OH position and glycosylate isoflavones (daidzein, formononetin, and genistein) at the 7-OH or 4'-OH position, exhibiting no enzyme activity to flavonones (liquiritigenin and narigenin) in vitro. The identification of PtUGT8 may provide a useful enzyme catalyst for efficient biotransformation of isoflavones and other natural products for food or pharmacological applications.
Cloning, Molecular ; Genistein ; Glucosyltransferases/metabolism* ; Isoflavones/pharmacology* ; Pueraria/chemistry*

OBJECTIVETo investigate the metabolism-based interaction between ipriflavone(IP) and other drugs in vitro.

METHODSIP and propafenone or estradiol, or other drugs were co-incubated with rat liver microsome induced by beta-naphthoflavone. The concentrations of residual substrate in microsomal incubates were determined by HPLC.

RESULTThe metabolism of propafenone (10 microg/ml) incubated with 2 microg/ml or 50 microg/ml IP was markedly inhibited compared with control group(P<0.01). While the metabolism of 10 microg/ml of estradiol incubated with 10 microg/ml and 100 microg/ml IP showed no marked inhibition. When IP (20 microg/ml) was incubated with 0.5 microg/ml propranolol, 0.5 microg/ml propafenone or 5.0 microg/ml estradiol respectively the metabolic activity of IP was markedly inhibited compared with control group (P<0.05 P<0.05 P<0.02 respectively).

CONCLUSIONThe results indicate that there is an interaction between ipriflavone and propafenone during in vitro metabolism, and there might also be clinical significance.

Animals ; Drug Interactions ; Estradiol ; metabolism ; Female ; Isoflavones ; metabolism ; Propafenone ; metabolism ; Propranolol ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the corneal penetration of PAMAM dendrimers-coated puerarin liposomes in rabbits.

METHODEvaluated PAMAM (G2, G3) dendrimers-coated puerarin liposomes were prepared and the in vitro transcorneal penetration were compared to puerarin drop solution and uncoated liposomes. The effect of different proportion of PAMAM to phospholipids in formulation on corneal penetration and the penetration parameters were investigated.

RESULTThe steady state fluxes and permeability coefficients of puerarin by PAMAM G2 (1.0%) and PAMAM G3 (0.5%) coated puerarin liposomes were greater than that by puerarin drop solution and uncoated liposomess (P < 0.01), meanwhile the PAMAM G2 (1.0%) and PAMAM G3 (0.5%) coated liposomes were better than other ratios of coated liposomes for improvement of corneal penetration (P < 0.01).

CONCLUSIONThe PAMAM coated liposomes is able to enhance the corneal penetration of puerarin and promising as an ocular drug carriers.

Animals ; Cornea ; metabolism ; Dendrimers ; chemistry ; metabolism ; In Vitro Techniques ; Isoflavones ; chemistry ; Liposomes ; chemistry ; metabolism ; Rabbits

The study aims to screen the ability of the bacteria to metabolize ononin and assess the effect of ononin on the intestinal bacteria. Fresh human fecal sample was obtained from a healthy volunteer, diluted serially in sterile water and sixty-nine different bacterial colonies were picked out ultimately. UPLC-Q-TOF/MS with automated data analysis software (MetaboLynx) was applied to fast analysis of ononin metabolites. Furthermore, an E(max) precision microplate reader was employed to determine the growth situation of Enterococcous sp., Enterobacter sp., Lactobacilli sp., and Bifidobacteria sp. Results indicated that hydrogenation, demethylation, hydroxylation and deglycosylation were the major metabolic pathways of ononin by human intestinal bacteria in vitro. Ononin can inhibit the growth of pathogen such as Enterococcus sp., Enterobacter sp. and can promote the growth of probiotics such as Bifidobacteria sp. and Lactobacilli sp. This study suggested that intestinal bacteria have the metabolic effects of ononin and the biotransformation was completed by different bacteria. And ononin can affect the balance of intestinal flora and the degree of influence varies depending on the bacterial species and the concentration of ononin.
Bacteria ; metabolism ; Biotransformation ; Feces ; microbiology ; Glucosides ; metabolism ; Humans ; Intestines ; microbiology ; Isoflavones ; metabolism ; Metabolic Networks and Pathways

The gelatinization process of the starch is replaced by unpolluted steam-pretreatment on the base of the Radix Puerariae rich in fiber and isoflavones. The production of ethanol and isoflavones by simultaneous saccharification and solid state fermentation (SSF) of steam-pretreatment Radix Puerariae is presented. The optimal technological conditions were obtained: Radix Puerariae being steam-pretreated at a saturated vapor pressure of 0.8 MPa for 3.5 min, glucoamylase(65 u/g), cellulase(1.5 u/g), 0.1%(NH4)2SO4, 0.1%KH2PO4 and activated yeasts being added in, and fermentation at 35-37 degrees C for 60 h. Under these conditions, the yield of ethanol and isoflavones from 100 g Radix Pureriae (dry basis) were 27.47 g and 4.43 g, respectively, the starch utilization rate was 95%. In comparison with the traditional fermentation technology, the simultaneous saccharification and SSF of steam-pretreatment Radix Puerariae is clean and energy-saving. It provides new way of the production of ethanol from the non-food starch material, and worthwhile to be explored and implemented in industry.
Cellulase ; metabolism ; Ethanol ; metabolism ; Fermentation ; physiology ; Glucan 1,4-alpha-Glucosidase ; metabolism ; Isoflavones ; biosynthesis ; Pueraria ; metabolism ; Steam ; Yeasts ; metabolism

A high activity isoflavone-glucosidase, which hydrolysis glycosides, was obtainde using liquid fermentation from Absidia sp. R strain. The isoflavone-glucosidase was purified 11 folds with yielding rate of 10.9% after ammonium sulfate precipitation and DEAE-Cellocuse (DE-52) ion exchange chromatography. SDS-PAGE results showed that the molecular weight is 53kD. And the optimum temperature, the optimum pH, Km and pI of the enzyme are 50 deegrees C, 5.0, 1.3 x 10(-2) mol/L and 3.2, respectively. The isoflavone-glucosidase is also rather stable under 60 degrees C and in pH range from 5.0 to 7.0. The enzyme can be activated by Co2+ and Ca2+, and be inhibited by Ag+ and Cu2+.
Absidia ; enzymology ; Glucosidases ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Isoflavones ; metabolism ; Temperature

PURPOSE: It has been postulated that soybean isoflavones act as inhibitory factors in several cancers. Recently, various in vitro and in vivo experimental studies have demonstrated that these isoflavones inhibit prostate cancer. Therefore, we investigated whether soybean isoflavones influenced the development of prostate cancer by comparing the levels of circulating isoflavones between prostate cancer patients and controls. MATERIALS AND METHODS: The serum levels of genistein, daidzein and equol were determined using reverse-phase, high-performance liquid chromatography-multiple reaction ion monitoring mass spectrometry(HPLC-MS) and compared in 122 experimental subjects(61 prostate cancer patients and 61 cancer-free controls) from 6 hospitals. RESULTS: The serum concentrations of genistein, daidzein and equol in the patients were 130.7+/-181.4ng/ml, 53.6+/-69.3ng/ml and 11.37+/-43.4ng/ml, with control values of 95.6+/-95.2ng/ml, 55.2+/-121.8ng/ml and 23.2+/-34.5ng/ml, respectively. There was no statistical difference between the 2 groups. Daidzein non-metabolizers who were unable to degrade daidzein into equol were compared between the patients and the controls, and were found to be significantly more common in the patient group (p=0.001, OR=3.44, 95% CI=1.6243-7.2855). However, the equol/daidzein ratio was significantly lower in the patients than in the controls(p=0.0072). No association between age, stage, Gleason score or isoflavone concentrations was found. CONCLUSIONS: These results suggest that the capability to produce equol (i.e., the mechanism for the metabolism of daidzein into equol) is closely involved in the lower incidence of prostate cancer, and that a diet based on soybean isoflavones would be useful in preventing prostate cancer.
Diet ; Equol ; Genistein ; Humans ; Incidence ; Isoflavones ; Metabolism ; Neoplasm Grading ; Prostate* ; Prostatic Neoplasms* ; Soybeans*

OBJECTIVETo establish the model of microdialysis, and study the ocular pharmacokinetics of puerarin in anesthetic rabbits.

METHODImplanted the probe into anterior chamber of anesthetic rabbit by surgery. After balanced for 2 h, 1% puerarin eye drop (100 microL) was applied into the cul-de-sac with micropipette. Immediately the dialysate was collected at different time and detected by HPLC with the detection wavelength of 249 nm. The mobile phase was methanol and 0.1% citric acid solution (30:70); the flow rate was 1.0 mL x min(-1).

RESULTAfter the administration, puerarin can be absorbed into aqueous humor quickly. The peak concentration of puerarin appeared at about 1 h and then reduced gradually. The peak concentration(C(max)) is (2.52 +/- 0.31) mg x L(-1). The other lower peak was shown at 3.5 h during the eliminate phase. This might be attributed to the inhibition of aqueous humor production by the puerarin and resulted in a high drug concentration. The area under concentration-time curve (AUC(0-t)) is (5.04 +/- 0.21) mg x h x L(-1) and the eliminate half life (t1/2) is (0.38 +/- 0.13) h.

CONCLUSIONThe microdialysis technique can be used to detect the ocular pharmacokinetics of puerarin, and support the valuable pharmacokinetics parameter for the clinical applications of puerarin eye drop.

Anesthesia ; Animals ; Eye ; metabolism ; Female ; Isoflavones ; pharmacokinetics ; Male ; Microdialysis ; methods ; Ophthalmic Solutions ; Rabbits

The overall purpose of this study was to investigate the effects of level of isoflavones supplementation on bone metabolism in growing female rats. The effects of level of isoflavones supplementation on bone mineral density (BMD) and bone mineral content (BMC) were inspected in this study. Forty-five rats divided into three groups: Casein, 1/2IF, IF. The serum and urine concentrations of calcium and phosphorus were determined. BMD and BMC were estimated by using PIXImus (GE Lunar Co, Wisconsin.) in spine and femur on 3, 6, 9 weeks after feeding. This study of results were as follows: The isoflavones supplementation level did not affect weight gain, mean food intake and food efficiency ratio. The serum concentration of calcium, phosphorus were not significantly different by different level of isoflavones supplementation. The urinary calcium and phosphorus excretion were not significantly different, too. Spine and femur BMD, BMC were not significantly increased by different level of isoflavones supplementation on 3 and 6 weeks after feeding. Spine BMD and spine BMC per weight, femur BMC per weight were significantly increased in the groups 1/2IF and IF at the ninth week after feeding, but there was no significant difference by different level of isoflavones supplementation. Spine BMD per weight and femur BMD per weight were significantly higher in the group of IF than in the group of Casein and 1/2IF at the ninth week after feeding. These results suggest that the group of IF with rich isoflavones supplementation was effective to the increase of BMD spine and femur in growing female rats, respectively.
Animals ; Bone Density* ; Calcium ; Caseins ; Eating ; Female* ; Femur ; Humans ; Isoflavones* ; Metabolism ; Phosphorus ; Rats* ; Spine ; Weight Gain

OBJECTIVES: Menopause is associated with adverse metabolic changes in plasma lipoprotein and inflammation markers. Estrogens have beneficial effects on lipid metabolism and inflammation. Isoflavones (ISO) have structurally similar to estradiol. Our objective was analize the effect of soy-ISO on serum lipid and inflammatory markers (sP-selectin and sCD40L) in postmenopausal women. METHODS: A 12-week randomized, double-blind, placebo-controlled intervention with soy-ISO (50 mg, twice daily) was conducted in 35 healthy postmenopausal women (55–72 years old). The women were divided in 2 groups: 20 were allocated to soy-ISO, and 15 to a placebo group. RESULTS: The changes of total cholesterol (TC), triglycerides, low-density lipoproteins-cholesterol (LDL-C), high-density lipoprotein-cholesterol, Apo-A1, sP-selectin and sCD40L in 2 groups before and after 12-week treatment showed no statistical significance. In subgroup analysis, soy-ISO supplementation significantly decreased the levels of TC, LDL-C and sCD40L in women under 65 years old, and with null effects on serum lipid and inflammation markers in women over 65 years old. CONCLUSIONS: Soy-ISO did not significantly favorable effects on the lipid profile and inflammatory markers in postmenopausal women. However, in women under 65 years of age, soy-ISO significantly decreased the TC, LDL-C and sCD40L, whereas, no effects on lipid profile and inflammation markers in women over 65 years old were observed.
Cholesterol ; Dyslipidemias ; Estradiol ; Estrogens ; Female ; Humans ; Inflammation ; Isoflavones ; Lipid Metabolism ; Lipoproteins ; Menopause ; Plasma ; Postmenopause ; Triglycerides