Experimental Study On Jiaweisinisan Against Liver Fibrosis

1 ObjectiveHepatic fibrosis,which changes slow-moving,is a common pathological process of varied chronic liver disease to cirrhosis.Now,there isn't any effective method to cure the diseases leading to hepatic fibrosis,so it appears very important to prevent or delay the development of hepatic fibrosis,and it also is a focal point of study.Based on experiences of treating chronic hepatitis and liver fibrosis for many years,we make up a compound formula called Jiaweisinisan with Radix Salviae Miltiorrhizae,Radix Notoginseng and Carapac Trionycis added to Sini Powder for Soothing Liver and Regulating Qi which is a classical formula for TCM.This thesis has carried on related experiments to explore the pharmacodynamic action and mechanism t of Jiaweisinisan against liver fibrosis,which will provide the theory evidence for the herbs.2 Experimental contentsThis part is made up of three experiments:2.1 Study of Jiaweisinisan in vivo70 SD rats were used for this experiment and were divided into six groups at random.Except the normal group,the rest were injected with 0.5%Dimethylnirtosamine (DMN),three times a week,and were treated with Jiaweisinisan in experimental group,colchicine tablets in positive controlled group.After the four weeks' treatment with Jiaweisinisan,we detected the rats' ALT,ALP,GGT,TP,ALB,HA and C-â…£from their blood serum.Hydroxyproline(HyP)was also detected from liver.At the same time,the change of the rats' histopathology was observed.2.2 Study of Jiaweisinisan on the growth of HSC-T6 in vitroHSC-T6 was used as the model in vitro.The stock solution of Jiaweisinisan was diluted by multiple proportion into 8 densities,and affected HSC-T6.After 72 hours,MTT was added to every hole by 0.1ml.Then every hole's absorbance was determined by a automatic machine on 570nm 4 hours later.Referring to a formulae,TC₅₀was calculated to analyze the effect of Jiaweisinisan on the growth of HSC-T6.2.3 Study of Jiaweisinisan on the mRNA of TGF-β₁ in HSC-T6 in vitroThis part was based on the above experiment.HSC-T6 in vitro was affected by 1/2 TC50 and 1/4 TC50 of Jiaweisinisan.Then under the condition of RNA-free, the mRNA of TGF-β₁ was gotten out step by step.The result was followed by RT-PCR.Then wen can analyze the effect of Jiaweisinisan on the mRNA of TGF-β₁3 ResultsThe results of experiment 1:for the rat models of liver fibrosis induced by Dimethylnirtosamine(DMN)and treated with Jiaweisinisan,the survival rats of the normal group,the model group,the positive controlled group and the experimental groups are 100%,71.43%,75%,91.67%,100%,80%.And the serum levels of ALT,ALP,GGT,HA,C-â…£and the HyP from hepatic tissue decrease significantly.At the same time,the levels of TP and ALB increase.Accordingly,the pathologic lesions of hepatic fibrosis also significantly improve in post-treatment (P＜0.01).In the experiment 2,the formula Jiaweisinisan of 100mg/mLand 50mg/mL have an inhibitory action on the growth of HSC-T6.The TC₅₀is 51.51mg/mL.The experiment 3 appears that Jiaweisinisan can restrain the mRNA of TGF-β₁ in HSC-T6 in vitro notably(P＜0.05),and promote the apoptosises of HSC-T6.4 ConclusionsAll the experiments have proved that Jiaweisinisan has an obvious effect on liver fibrosis.It could restrain the expressions of correlated cytokines and promote the apoptosises of HSC-T6,which may be the mechanism against liver fibrosis.These results prognosticate the good outlook of Jiaweisinisan against liver fibrosis and provide the evidence on pharmacodynamic action for clinical application.