The Pharmacodynamics Study On Anti-Experimental Liver Injury And Anti-HBV Of Total Paeony Glycoside(TPG)

The resource of Chinese herbs is abundant, some of herbs have protectiveeffects on CHB, especially single herb, which is to be developed as the idealmedicine. During long-term experimental and clinical studies, we elected TPGdistilling from the Peony Root through repeatedly filtrated and optimized,then a new potential anti-HBV and anti-liver indury herb is being developed.Objective1. Animal models were used whose livers were damaged by D-GalN and Con Ato study the hepatoprotective effect of TGP as well as its hepatic tran-saminase lowering capability.2. Study the anti-HBV activity of TGP in vitro and in vivo.The step of experiments1. The TPG's protective effect of the liver injury caused by D-GalN.2. The TPG's protective effect of the liver injury caused by Con A.3. Experimental study of the anti-DHBV effect of the TPG in vivo.4. Experimental study of the anti-HBV effect of the TPG in vitro.Methods1. Sixty NIH mice were randomly divided into 5 groups normal control group(groupA), model group(groupB), high dosage group of TPG (groupC), low dosagegroup of TPG(g-roupD) and bifendate group (groupE). Mice in group C, groupD andgroupE were administered with drugs by gastric perfusion for 7 days once dailyconsecutively. One hour after the last administration of the drugs, exceptsthose in group A, mice in other groups were injected with D-GaIN 800mg/Kg intoits abdominal cavity. Blood samples for detecting ALT,AST,superoxide dis-mutase(SOD) activity and malondiadehyde(MDA) concentration were collectedat the 16 hours after D-GaIN injected. Histopathological examintion was also performed for liver tissue.2. Fifty NIH mice were randomly divided into 5 groups: normal control group(groupA), model group (group B), high dosage group of TPG (dose of 20mg/kg,group C), low dosage group of TPG ((dose of 10mg/kg, group D)and bifendategroup (group E). Except those in group A, mice in other groups were injectedwith Con A (dose of 20mg/kg) via the tail vein at the first day of experiment.Group C, group D and Group E were administered by gastric perfusion respect-ivity at the dose of TPG 20mg/kg, 10mg/kg and bifendate 150mg/kg. All the drugswere given once daily for 3 days continuously. At the last time, when drugswere administered for 4 hours, mice were injected with Con A once again atthe same dosage. Blood samples for aminotransferase (ALT),AST activity andtumor necrosis factor alpha (TNF-α) content measurement were obtained at the8th hour after Con A injected. Histopathological examination was also per-formed for liver tissue.3. The 1 day old congenitally infected duck was divided to 4 groups randomly,every group was 6-7 ducks differentenly. Namely DHBV control group, ACV group,high dosage group of TPG, low dosage group of TPG. The intragastric adminis-tration dose of TPG were repectivity 20mg/kg. lOmg/kg, ACV was 100 mg/kg. Thecontrol group was not treated. The serum was collected in the time points ofT0,T5,T10 and P3 in accordance with the time point of drug intragastricadministration. The quantity of DHBV in these specimens was evaluated bydot-blot assay.4. rake the 2.2.15 cell line as cell model, the TPG were divided into 7 dosagegroups. After 6 days drug administration, The HBeAg and HBeAg in the supern-atant was tested with ELLSA assay. The toxicity of drug to the cell wasevaluated with the MTT assay. The TI was computed.Results1. The high dosage Total TPG significantly lowered serum ALT,AST and MDAIn the liver tissue, enhanced SOD in the liver tissue and markedly amelioratedthe liver damage(p＜0.05).2. ALT,AST activity and TNF-αcontent in group C was obviously lower to thatof group B(P＜0.01), but no significan difference comparing with groupE (p＜0.05),hepatichistop-athological changes were alleviated in group C.3. The titer of blood DHBV was significantly drop down after 10 days drugadministration (P＜0.01). The titer of blood DHBV was ascended in 3 days Post drug administration. Contrasted to the DHBV control group, P＞0.05. This indi-cate that the inhibit effect of the TPG to the DHBV was occurred after 10 daysdrug administration. But the DHBV can replapse after 3 days stopping of drugadministration.4. It was observed that the semi-toxic concentration(TC₅₀)of Total paeonyGlycoside on 2.2.15 cell strain was 0.17g/L 6 days after action on the 2.2.15cell strain and the half inhibitory concentration (IC₅₀)on HbsAg was 0.15g/Land that on HBeAg was 0.06s/L, The treatment index on HBsAg positives was 1.12and 2.75 on HBeAg positives.Conclusion1. The TPG can protect against acute liver injury induced by D—GalN in mice,the mechanism of which maybe related to its anti—OFR injury.2. The TPG has good protective effect on liver injury in mice induced byCon A by suppressing the activation of the T-lymphocytes and reducing therelease of TNF-α.3. The TPG has anti-DHBV effect in vivo. But in the congenitally infectedduck animal model, The DHBV may replapse after stopping of drug administration.4. Total paeony glycoside possesses inhibitory effect on the secreion of HBsAglowly effectively and toxicly, and on the secretion of HBeAg effectively andlowly toxicly in vitro culture cells respectively.