Polygonflavanol B(1), a new flavonostilbene glycoside, was isolated from the roots of Polygonum multiforum(Polygonaceae) by various column chromatography methods including macroporous resin HP-20, silica gel, Sephadex LH-20, and preparative HPLC. The structure with absolute configuration of the new compound was identified by its physicochemical properties, spectroscopic data, ECD calculation, and chemical method.
Fallopia multiflora/chemistry* ; Flavonols/isolation & purification* ; Glycosides/isolation & purification* ; Plant Roots/chemistry* ; Stilbenes/isolation & purification*

A reasonable and practicable quality standard was developed for mori liquid extract from different sources by TLC, HPLC and fingerprint technology. In TLC method, the compounds were separated on polyamide film using glacial acetic acid-water (1: 3) as mobile phase at a UV wavelength of 365 nm. All qualified samples had the spots of the same color as the control herb and substance. The RP-HPLC method was used to determine the content of mulberroside A with mobile phase of methanol-water (25: 75) at a wave-length of 326 nm. The mulberroside A was in good linear with a regression equation of Y = 46.965X (r = 0.999 6) in the range of 4.6 - 228 mg x L(-1). In 14 batches of samples, the mulberroside A in 4 batches of them was less than 0.5 g x L(-1), and was more than 2.0 g x L(-1) in the other batches. It was suggested that the content limit of mulberroside A should be no less than 1.5 g x L(-1). The HPLC fingerprints were evaluated by the similarities. It has found that the similarities of different mori liquid extracts were very low and the chemical diversity of mori cortex was the major factor of similarity. Moreover, the process impact was minimal. Thus the fingerprint was not included in this quality standard.
China ; Chromatography, High Pressure Liquid ; Disaccharides ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Morus ; chemistry ; Quality Control ; Stilbenes ; chemistry ; isolation & purification

OBJECTIVETo study major antioxidant active compounds in Mori Ramulus.

METHODThe combination of on-line HPLC method and liquid chromatography-mass spectrometry was used to identify major antioxidant compounds and their content was determined by HPLC-DAD (detection wavelength of 320 nm).

RESULTOxyresveratrol was proved to be the major antioxidant compound in Mori Ramulus ethanolic extracts. Using the HPLC-DAD quantitative determination, Mori Ramulus oxyresveratrol had a good linear range of 14-1 260 mg x L(-1) (r = 0.999 96). The average recovery was 98.2% with RSD of 1.2%. This method is simple, rapid, accurate and sensitive. There was rich oxyresveratrol in Mori Ramulus and the content was significantly different according to the mulberry varieties.

CONCLUSIONThe on-line HPLC-MS-DPPH method is applicable for the determination of antioxidant compounds in Mori Ramulus.

Antioxidants ; analysis ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; toxicity ; Mass Spectrometry ; Medicine, Chinese Traditional ; Morus ; chemistry ; Plant Extracts ; analysis ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Reproducibility of Results ; Sensitivity and Specificity ; Stilbenes ; analysis ; chemistry ; isolation & purification ; Time Factors

OBJECTIVE: To synthesize 3, 5, 4' -trimethoxystilbene (TMS) by methylation of resveratrol (Res), a natural compound extracted from polygonum cuspidatum, to identify the chemical structure of TMS, to test its pharmacokinetics, and to determine the effects of TMS on the growth inhibition and apoptosis in pulmonary artery smooth muscle cells (PASMCs). METHODS: The chemical structure of TMS was analyzed by UV- and IR- absorption spectrometry, (1)H-NMR and (13)C-NMR spectroscopy and mass spectrometry. We measured the bioavailability, the characteristics of intestinal absorption, and the distribution of TMS in body and excretions of SD rats after oral administration of TMS. The acute toxicity of TMS in mice was tested. PASMCs were prepared from pulmonary artery of SD rats. The PASMCs were divided into 8 groups. Group of A (control) was cultured without TNF-α, TMS, or Res. Group of B (TNF-α) was cultured with 100 pg/mL TNF-α. Groups of C-E (low-high concentrations of TMS) were cultured with 100 pg/mL TNF-α and 5, 10, 20 μmol/L TMS, respectively. Groups of F-H (low-high concentrations of Res) were cultured with 100 pg/mL TNF-α and 50, 100, 200 μmol/L Res, respectively. The proliferation of PASMCs after treatment was determined by MTT assay. The apoptosis of PASMCs after treatment was determined by flow cytometry. RESULTS: The UV absorption map of TMS showed λmax(MeOH) at 318, 306.2, and 217.8 nm. Analysis of infrared spectrum of TMS showed IRvKBr max /cm at 2999, 2935, 2836, 1591, 1511 and 1456/cm. The (1)H-NMR map showed that the synthetic product contained three hydroxy groups, while (13)C-NMR map showed 17 carbon signals and some symmetrical structural fragments. Electospray ionization mass spectrometry of the productshowed m/z peaks corresponded to 271[M+H](+), 256[M+H-CH(3)](+) and 241[256-CH(3)](+); the implied relative molecular weight is 270 and the implied molecular formula is C17H18O3. These data confirm the product is 3,5,4' - trimethoxystilbene. The absolute bioavailability of TMS was 45.4%. TMS was well absorped in the upper small intestine; it was excreted in stool and bile and distributed into several tissues. The maximal tolerance dose (MTD) of TMS was 5.85 g/kg. MTT assay showed TMS inhibited the proliferation of PASMCs in a dose-dependent manner. The extent of growth inhibition in A-H groups were (4.07±2.12)%, (6.54±4.78)%, (9.35±4.26)%, (16.75±5.34)%, (23.74±7.07)%, (6.78±5.58) %, (8.81±5.16) %, and (17.81±6.03) %, respectively. Flow cytometry showed the extent of apoptosis in PASMCs (after being treated with TMS for 24 h) was significantly higher than that in PASMCs treated only with TNF-α. The apoptosis rates of A-H groups were (2.63±0.74)%, (3.54±0.81)%, (5.77±4.62)%, (11.68±5.35)%, (18.79±4.15)%, (4.11±3.59)%, (6.33±4.8) %, and (12.47±5.06)%, respectively. CONCLUSION We have confirmed our synthetic product as 3,5,4'-trimethoxystilbene (TMS), with the molecular formula of C17H18O3 and appropriate molecular weight and absorbption and NMR spectra. The bioavailability of TMS was to 45%. It strongly inhibits the proliferation of PASMCs in a dose-dependent manner and induces apoptosis of PASMCs.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; Cells, Cultured ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Pulmonary Artery ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Resveratrol ; Stilbenes ; chemical synthesis ; chemistry ; isolation & purification ; pharmacokinetics ; pharmacology

Resveratrol, isorhapontigenin and pinosylvin, isolated from Gnetum parvifolium, and their analogues have been synthesized and tested for their inhibitory activity of HIV-1. Natural product 12a and analogues (12d, 12e, 12g) display significant inhibitory activity of HIV-1 replication. Among them, compound 12d (trans-3, 4, 5, 4'-tetrahydroxystilbene) exhibits the most potent anti-HIV-1 activity with an IC50 value of 1.84 micromol x L(-1).
Anti-HIV Agents ; isolation & purification ; pharmacology ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gnetum ; chemistry ; HIV-1 ; drug effects ; physiology ; Inhibitory Concentration 50 ; Leukocytes, Mononuclear ; cytology ; virology ; Molecular Structure ; Plants, Medicinal ; chemistry ; Stilbenes ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo optimize the ultrahigh pressure extraction (UPE) process of polydatin and resveratrol from Polygonum cuspidatum by using uniform design.

METHODOn the basis of single factor screening, the uniform design was adopted for getting the optimal technique parameters. The optimum result of UPE was compared with conventional extractions.

RESULTThe optimal conditions of UPE for polydatin and resveratrol were that the solvent was 55% ethanol, the ratio of solvent to material( mL: g) was 30, the extraction pressure was 170 MPa, and the extraction time was 120 second. With this extracting process, the extraction yield of polydatin and resveratrol were 14.29 and 2.53 mg x g(-1), respectively. The extraction yield of polydatin was 46.1% higher than the heat reflux extraction, 6.4% higher than the ultrasonic extraction and 28.5% higher than the microwave extraction, while the yield of resveratrol was 67.5% higher than the heat reflux extraction, 29.7% higher than the ultrasonic extraction and 24.6% higher than the microwave extraction, respectively.

CONCLUSIONAs a novel extraction technology for Chinese herbal medicine, the UPE procedure has higher extraction yield, lower extracting temperature, shorter extacting time and less power consumption. The UPE has provided a brand-new method for extraction of polydatin and resveratrol from P. cuspidatum.

Calibration ; Chemical Fractionation ; methods ; Fallopia japonica ; chemistry ; Glucosides ; analysis ; isolation & purification ; Pressure ; Reproducibility of Results ; Solvents ; chemistry ; Stilbenes ; analysis ; isolation & purification ; Time Factors

AIMTo observe the changes of MMP-2, 9 level on atherosclerosis in experimental rats by treatment of 2,3,4',5-tetrahydroxystilbene-2-0-beta-D glucoside (TSG) and to investigate the mechanism of TSG in stabilizing plaque and anti-atherosclerosis.

METHODSThe atherosclerosis model of rat was made by feeding high grease food and injecting VitD3. Sixty male SD rats were randomly divided into six groups: control, Simvastatin, model and TSG 120 mg x kg(-1) x d(-1), TSG 60 mg x kg(-1) x d(-1) and TSG 30 mg x kg(-1) x d(-1). After 12 weeks, several aorta were randomly tested, model and TSG 120 mg x kg(-1) x d(-1), TSG 60 mg kg(-1) x d(-1) and TSG 30 mg x kg(-1) x d(-1). After 12 weeks, several aorta were randomly tested, model made was successful when we found plaque. And after six weeks treating, the mRNA expressions of MMP-2 and MMP-9 were measured by RT-PCR. The activities of MMP-2 and MMP-9 were measured by Western blot. The levels of CRP, IL-6 and TNF-alpha in serum were measured in biochemical method.

RESULTSData of the study demonstrated that the level of TNF-alpha, IL-6, CRP, MMP-2 and MMP-9 were remarkably decreased by TSG60, 120 mg x kg(-1) x d(-1) groups, which showed a dose-dependent effect.

CONCLUSIONTSG has the effect of anti-atherogenic and stabilizing plaque on the experimental rats with atherosclerosis, which are induced by the high cholesterol feeding and VitD3 injecting. The effect of TSG seems to be closely involved in regulating the expressions of MMP-2 and MMP-9, and inhibiting inflammation.

Animals ; Atherosclerosis ; drug therapy ; metabolism ; C-Reactive Protein ; metabolism ; Glucosides ; isolation & purification ; therapeutic use ; Interferon-alpha ; blood ; Interleukin-6 ; blood ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Polygonum ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; isolation & purification ; therapeutic use

Cajanus cajan L. is a natural plant, which contains a lot of potential active components. In the present study, we identified the effects of the stilbene extract from Cajanus cajan L. (sECC) on hepatic cholesterol metabolism in diet-induced (for 4 weeks) hyperlipidemic Kunming mice. All experimental mice were divided into 5 groups: control group, high lipid model group, sECC-treated with 200 or 100 mg kg(-1), and simvastatin (Sim, 12 mg kg(-1)) treated group. The mice were fed with fat and cholesterol-enriched chow except control mice that were fed with standard diet. The effects of sECC were investigated by monitoring serum and liver lipid profile (i. e. cholesterol homeostasis) in mice. To further explore the mechanism of sECC, hepatic cholesterol 7alpha-hydroxylase (CYP7A1) and low density lipoprotein (LDL) receptor expressions in cholesterol homeostasis were analyzed by reverse transcription PCR. After 4 weeks pretreatment, the mice in the high lipid model group showed markedly higher serum and hepatic lipid contents than control group (P< 0.01). Compared with high lipid model group, the increased serum and hepatic lipid contents were markedly attenuated by sECC (200 mg kg(-1)), the serum and hepatic total cholesterol were reduced by 31.5% and 22.7% (P<0.05), respectively. The triglyceride contents of serum and liver were also lowered by 23.0% and 14.4%, respectively. At the same times, serum LDL cholesterol decreased by 53.0% (P<0.01). The mRNA expressions of hepatic CYP7A1 and LDL-receptor were significantly enhanced in the mice administered with sECC (200 mg kg(-1)), whereas those expressions were suppressed by the fat and cholesterol-enriched diet. These data indicate that sECC reduces the atherogenic properties of dietary cholesterol in mice. It is indicated that expression enhancement of hepatic LDL-receptor and cholesterol 7alpha-hydroxylase may be responsible for the hypercholesterolemic effect.
Animals ; Anticholesteremic Agents ; isolation & purification ; pharmacology ; Body Weight ; drug effects ; Cajanus ; chemistry ; Cholesterol ; blood ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; biosynthesis ; genetics ; Cholesterol, LDL ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Expression Regulation ; Hypercholesterolemia ; blood ; genetics ; metabolism ; pathology ; Liver ; metabolism ; pathology ; Male ; Mice ; Organ Size ; drug effects ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Stilbenes ; isolation & purification ; pharmacology ; Triglycerides ; blood ; metabolism

The investigation on the stem bark of Morus wittiorum was carried out to find its chemical constituents possessing anti-oxidative activity. The isolation and purification were performed by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography and so on. Based on the spectral analysis such as NMR, MS, etc., seven 2-arylbenzofuran derivatives were identified as wittifuran D (1), wittifuran E (2), moracin C (3), moracin M (4), moracin P (5), 2-(3,5-dihydroxyphenyl)-5,6-dihydroxybenzofuran (6) and mulberroside C (7). Compounds 1-7 were isolated from this plant for the first time. Among them, 1 and 2 were new compounds. Compounds 3-7 were used to assay antioxidant activity, the inhibitory ratios of compounds 3, 4, 6, at a concentration of 1 x 10(-5) mol x L(-1), to malondialdehyde (MDA) produced during microsomal lipid peroxidation induced by ferrous-cysteine were 73%, 69% and 89% respectively.
Antioxidants ; isolation & purification ; pharmacology ; Benzofurans ; chemistry ; isolation & purification ; pharmacology ; Lipid Peroxidation ; Malondialdehyde ; chemistry ; Molecular Structure ; Morus ; chemistry ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry ; Stilbenes ; chemistry ; isolation & purification ; pharmacology

This study was intended to look for anti-HIV chemical constituents of aerial parts of Caragana rosea Turcz. Column chromatographic technique was used for the isolation and purification of constituents of Caragana rosea under the guide of anti-HIV assay. The structures were established on the basis of physical and chemical properties and spectroscopic data. Five compounds were obtained from the EtOAc fraction of aerial parts of Caragana rosea and identified as myricetin (1), mearnsetin (2), p-hydroxy cinnamic acid (3), cararosinol A (4) and cararosinol B (5). At the same time, one possible transformation route between cararosinol B and kobophenol A, another resveratrol tetramer isolated from this plant previously, was proposed. Compounds 4, 5 are new resveratrol tetramers, compounds 1 -3 were isolated from this plant for the first time. All compounds showed no activities in an in vitro assay against HIV-1.
Anti-HIV Agents ; chemistry ; isolation & purification ; pharmacology ; Benzofurans ; chemistry ; isolation & purification ; pharmacology ; Caragana ; chemistry ; Coumaric Acids ; chemistry ; isolation & purification ; pharmacology ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; HIV-1 ; drug effects ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Propionates ; Stilbenes ; chemistry ; isolation & purification ; pharmacology