ABSTRACT
OBJECTIVE：To establish the method for content determination of 6 components in Fuzheng guben granules ，such as 2，3，5，4′-tetrahydroxystilbene glucoside ，baicalin，icariin，scutellarin，baicalein and wogonin. METHODS ：HPLC method was adopted. The determination was performed on Dikma Diamonsil C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid aqueous solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 275 nm （0-8 min），320 nm（8-9 min）and 275 nm（9-33 min）. The column temperature was set at 25 ℃，and sample size was 10 μL. With baicalin as reference material ，the relative corr ection factors （fk/s） of other five components were calculated by multi-point correction method and slope correction method ；the retention time difference method was used to locate the chromatographic peaks ； the calculation values obtained by above 2 QAMS were compared with measured values of external standard method. RESULTS ： The linear range of 2，3，5，4′-tetrahydroxystilbene glucoside ，baicalin，icariin，scutellarin，baicalein and wogonin were 0.053-2.12， 0.163-6.52，0.059-2.36，0.021 6-0.864，0.03-1.2，0.021-0.84 μg（r＞0.999），respectively. RSDs of precision ，stability（12 h）and reproducibility tests were all lower than 3％. Average recoveries were 98.72％-99.82％（RSDs were 0.89％-1.24％，n＝9）. Using baicalin as reference material ，fk/s of multi-point correction method for 2，3，5，4′-tetrahydroxystilbene glucoside ，icariin，scutellarin， baicalein and wogonin were 1.172，0.528，1.479，1.820 and 2.534，respectively；fk/s of slope correction method were 1.234， 0.550，1.559，1.939，2.664. RSDs of 6 components in 10 batches of Fuzheng guben granules by 3 methods were 0.29％-2.77％ （n＝10），respectively. Pearson correlation coefficient was not lower than 0.999 9（P＜0.001）in measured values between QAMS and external standard method. CONCLUSIONS ：QAMS method is established successfully for simultaneous determination of 6 components in Fuzheng guben granules.
ABSTRACT
Objective: Mitochondrial dysfunction is evident in the early stage of Alzheimer's disease (AD). Therefore development of drugs that protect mitochondrial function is a promising strategy for AD. The present work was to investigate the effects of 2, 3, 5, 4′-Tetrahydroxystilbene-2-O-β-d-glucosides (TSG) on a mitochondrial dysfunction cell model induced by sodium azide and elucidate the underlying mechanisms. Methods: Mitochondrial membrane potential (MMP) was detected by a fluorescence method. Cellular adenosine triphosphate (ATP) level was measured using a firefly luciferase-based kit. Reactive oxygen species (ROS) was detected using dichlorofluorescin diacetate (DCFH-DA). The expression levels of Bcl-2 and Bax were measured by Western blotting assay. Flow cytometry was utilized to measure apoptosis. Results: Pretreatment of TSG (25–200 μmol/L) for 24 h significantly elevated MMP and ATP content, reduced ROS level and Bax/Bcl-2 ratio, and inhibited apoptosis in SH-SY5Y cells exposed to sodium azide. Conclusion: These results suggest that TSG protects SH-SY5Y cells against sodium azide-induced mitochondrial dysfunction and apoptosis. These findings are helpful to understand the protective effect of TSG on mitochondria, which are involved in the early stage of AD.
ABSTRACT
OBJECTIVE：To analyze the metabolites of tetrahydroxystilbene glucoside （THSG）and speculate its metabolism pathway in rats. METHODS ：Male SD rats were randomly divided into plasma group （n＝3），urine group （n＝3），bile group （n＝3），and tissue group （n＝9）. Each group was given single dose of THSG 200 mg/kg intragastrically. Plasma samples 10，30 min and 1，1.5，2，4 h after medication ，the unrine 0-6 h after medication ，the bile 0-4 h after medication ，the tissue of heart ， liver，spleen，lung，kidney and stomach 30 min and 1，2 h after medication （3 at each time point ）were collected respectively.After precipitated with methanol ，the metabolites of samples were analyzed and identified by UHPLC-Q-Exactive Orbitrap MS and mass loss filtration （MDF）. Its metabolism pathway was speculated. RESULTS：In the blood ，urine，bile，heart，liver，spleen， lung，kidney，stomach samples ，6，7，11，1，5，1，3，4，4 metabolites were detected ，including two phase Ⅰ（hydrolysis， hydrogenation and hydroxylation ）metabolites，18 phase Ⅱ（glucuronic acid binding and sulfation ）metabolites. There were 12 glucuronic acid binding products. CONCLUSIONS：Most of the metabolites of THSG are found in bile ，mainly glucuronic acid binding products of phase Ⅱ metabolite THSG ； main metabolic pathways involve glucose hydrolysis ， hydrogenation， hydroxylation，glucuronic acid binding and sulfation.
ABSTRACT
Objective To explore the influence of tetrahydroxystilbene glucoside ( TSG ) on the neuron proliferation in epileptic rats and the related mechanisms.Methods 54 healthy SD male rats were randomly divided into three groups: sham-operated group, epilepsy group and epilepsy with TSG treatment group.The epilepsy group was established by stereotactic brain trace injection with kainic acid ( KA ) . TSG solution ( 3 mg/kg ) was injected intraperitoneally at 6 hours after the epilepsy group established, and then q.d.for consecutive 42 days.The sham-operated group and epilepsy group were injected with normal saline.The influence of TSG on cell proliferation of rat hippocampal dentate gyrus BrdU-positive granular cells was observed by immunohistochemistry.Results Compare with the epilepsy group, the amount of glial fibrillary acid protein ( GFAP)-positive astrocytes was significantly reduced and dentate gyrus BrdU-positive cells were significantly increased in the TSG group ( P <0.01 ) .Conclusions Tetrahydroxystilbene glucoside ( TSG) promotes neurogenesis of neural stem cells and neurons, and inhibits the growth of hippocampal dentate gyrus astrocytes in epilepsy rats.
ABSTRACT
Aim: To investigate the effects of 2,3,5,4′-tetrahydroxystilbene -2-O-β-D-glucoside (TSG) on brain β-amyloid (Aβ) content, blood fat, and hemorheology in rat model induced by hypercholesterol. Methods The animal model of hypercholesterolemia was induced by feeding high cholesterol forage containing 1% cholesterol and 0.3% cholic acid for 10 weeks. TSG was given orally at doses of 30, 60 and 120 mg·kg-1 body wt/day for 10 weeks also. The content of β-amyloid was measured by immunohistochemistry and radioimmunoassay methods, the serum cholesterol and low density lipoprotein (LDL-C) were measured by automatic biochemistry analytical methods, and some indices of hemorheology were also observed. Results: The content of Aβ in hippocampus of rat model was increased after feeding high cholesterol forage for 10 weeks, and the serum cholesterol and low density lipoprotein level were obviously elevated as well, TSG shortened the content of Aβ in hippocampus, the serum cholesterol and low density lipoprotein level obviously. The indices of hemorheology showed that blood viscosity (ηb, high and low shear rate) and RBC adhesion index (AI) were increased in rats. TSG decreased blood viscosity (ηb, high and low shear rate) and lowed RBC adhesion index (AI) in rat model. Conclusion: TSG possesses obvious action of reducing the content of Ap in hippocampus, decreasing serum cholesterol and low density lipoprotein, promoting blood circulation and removing blood stasis. These actions may be related to the therapeutic mechanisms of AD.
ABSTRACT
@#ObjectiveTo observe the protective effect of tetrahydroxystilbene glucoside (TSG) on rats' hippocampal neuronal damage induced by glutamate (Glu) in the culture.MethodsHippocampus was isolated from newborn SD rats and dispersedly cultured in the medium for 9 days. Neurons were incubated with TSG (5—100μmol/L) for 24h, the cells were washed twice with Lock's solution without Mg2+,then Glu 500 μmol/L was added. Thirty min later, the reaction was terminated by washing the monolayer cells twice with the Lock's solution and then cultures were kept at 37℃ for 24h. Cell viability was measured by MTT method and cell membrane damage was determined by LDH leakage; with Fluo-3/AM as an intracellular calcium indicator and added into the bathing medium, fluorescent intensity of intracellular free calcium were observed through laser scanning confocal microscopy (LSCM).ResultsAfter the treatment with 5—100μmol/L TSG for 24h, the decrease of cell viability and the increase of LDH leakage caused by Glu was obviously resisted dose dependently. TSG inhibited increase of Ca2+ in cytoplasm, compared with model group.ConclusionTSG can significantly promote the cell viability and reduce the cell membrane damage in Glu treating hippocampal neurons. The neuroprotective activities of TGS is mediated by inhibiting Ca2+ overload in cytoplasm.
ABSTRACT
Aim To observe the effects of 2,3,4',5-tetrahydroxystilbene-2-O-?-D glucoside on expression of ICAM-1、VCAM-1 and VEGF in atherosclerotic rats,and approach the influence of TSG in atherogenesis.Methods Sixty SD rats were randomly divided into six groups(Control,Model,TSG 30、60、120 mg?kg-1?d-1 and Simvastatin 2 mg?kg-1?d-1 groups),given high-fat-diet for 12 weeks,the level of SOD and MDA then were determined.ICAM-1、VCAM-1 and VEGF protein were determined with Western blot assay,ICAM-1 mRNA and VCAM-1 mRNA were measured with RT-PCR,and VEGF expression was assessed by immunohisto-chemistry.Results In comparison with the model,the level of SOD increased markedly and the level of MDA decreased markedly after administering TSG;TSG could inhibit the expression of ICAM-1、VCAM-1 and VEGF in atherosclerotic arteries.Conclusions TSG could show its antiatherosclerotic effect by anti-oxydation and inhibiting the excretion of adhesion molecules.
ABSTRACT
Aim To investigate the effects of TSG on the content of nitric oxide and the expression of endo-thelium nitric oxide synthase in artery vessels of experimental atherosclerotic rats.Methods Sixty male rats were randomly divided into six groups.GroupⅠ,Normal control;GroupⅡ,Model control;Group Ⅲ,TSG low dose;Group Ⅳ,TSG middle dose;GroupⅤ,TSG high dose;group Ⅵ,Simvastatin.After 12 weeks,blood samples were collected from carotid arteries of rats,and the levels of NO in serum were measured.After that,the aortas were separated from the bodies and placed in-70℃,then the content of nitric oxide synthase was detected,and gene expression of eNOS and iNOSmRNA in artery vessels were respectively measured with RT-PCR method.STATA 8.0 software was adopted in date analysis..Results compared with those of the model group,the level of NO,the activity of NOS and the gene expression of eNOS were increased,and the gene expression of iNOS was reduced by simvastatin and TSG 30,60,120 mg?kg-1?d-1 in the high cholesterol-fed rats,which showed a dose-dependent effect.Conclusion TSG can enhance the expression of eNOS gene and reduce the expression of iNOS gene in aorta vessels of experimental atherosclerotic rats,which may be one of the anti-atherosclerosis mechanisms of TSG.
ABSTRACT
AIM: To investigate the preventive effect of tetrahydroxystilbene-glucoside (TSG) on experimental atherosclerosis rats. METHODS: Sixty one male rats were randomly divided into six groups. normal control; model control; TSG low dose(30 mg?kg-1?d-1);TSG middle dose(60 mg?kg-1?d-1);TSG high dose(120 mg?kg-1?d-1); simvastatin(2 mg?kg-1?d-1). The AS model of rats was made by feeding high grease food and injecting VitD3 .All the rats were fed for 12 weeks, blood samples were drawn from carotid artery of rats, the levels of TC, TG, HDL-C, LDL-C ,CRP,IL-6 and TNF-? in serum were measured with biochemical method. After blood samples were collected , the aorta samples were separated from the bodies, then they were placed 4% paraformaldehydea and were through Sudan Ⅳ and HE staining. STATA7.0 software was used to evaluate the differences between groups. RESULTS: Data of the study demonstrated that the levels of TC、TG、LDL-C、TNF-?、IL-6 and CRP were decreased remarkably, the level of HDL-C was increased by TSG 60, 120 mg?kg-1?d-1 group in the high cholesterol-fed rats, which showed a dose-dependent effect. The result of Sudan Ⅳ and HE staining suggested that the lipid deposits in aortic endothelium in TSG group were less than those in model group. CONCLUSION: TSG has preventive effect on the experimental atherosclerosis among the high cholesterol-fed rats. The anti-atherogenic effect of TSG seems to be closely related to regulating plasma lipid profile, and inhibiting inflammation.
ABSTRACT
Aim To observe the effects of 2,3,4′,5-tetrahydroxystilbene-2-0-?-D glucoside on expression of ICAM-1、VCAM-1 and VEGF excited by ox-LDL in U937 cell.Methods Differentiated U937 cells were cultured in vitro,excited by ox-LDL and intervened by different concentrations of TSG,then the level of SOD and MDA were determined.ICAM-1、VCAM-1 and VEGF protein were determined by Western blot and ICAM-1 mRNA and VCAM-1 mRNA were measured by reverse transcriptase-polymerase chain reaction(RT-PCR).Results In comparison with those of the control,the level of SOD remarkably decreased and the level of MDA remarkably increased in model group;the level of SOD increased markedly and the level of MDA decreased markedly in Simvastatin group and TSG 120 mg?L-1 group(P