Effects Of Antisense TIMP-1 Eukrayotic Expressing Plasmids On Experimental Liver Fibrosis In Vitro

Background: Hepatic fibrosis represents the final common pathological outcome for the major of chronic liver disease. There is a growing consensus that fibrosis belong to a reversible disease.Hepatic fibrosis is a pathological change that result from an imbalance between the deposition and degradation of ECM because of the continuous existence of insult and the repair of tissue. More deposition of ECM is more due to the reduce of degradation of ECM in the late stage besides of increase of synthesis. The process of hepatic fibrosis concerns complex cellular and molecular mechanism. Matrix metalloproteinase (MMPs)-tissue inhibitors of metalloproteinase (TIMPs) play an important role in liver fibrosis. MMP-1(human)/MMP-13(rat) may degrade typeâ… ,â…¢collagen which are the major component of ECM. TIMP-1 may inhibit the activity of MMPs. In the normal liver, the two factors keep in equilibrium state. The imbalance between the activities of the two factors by pathogeny contributes to ECM deposition in liver, which causes the hepatic fibrosis.HSC undergoes a phenotype transformation to a proliferating myofibroblast, which is the important access to hepatic fibrosis. During the development of fibrosis, MFB can transform to fibrocyte, which secret a mass of ECM. The active process of culture HSC in vitro can mimic the liver fibrosis process in vivo. The actived HSC increases synthesis of TIMP-1. TIMP-1 inhibits the activity of MMPs and the degradation of ECM, which contributes to hepatic fibrosis. HSC plays a prominent and central role in liver fibrosis and become the hotspot of research all over the world. Our present experiment was performed to isolate and culture HSC and transfer antisense TIMP-1 eukaryotic expressing plasmid into activated HSC in vitro. We aimed to test the effect of this exogenous plasmid and the possibility and effectiveness of gene therapy, which provided novel method to chronic fibrosis.Methods:1 HSC was extracted from normal anaesthetized Wistar rat by perfusing Collagenase and pronase E and purifying by density gradient centrifugation, were identified by immunohistochemistry.2 HSC was selected by growing in DMEM containing 0-600ug/ml G418. The concentration 450ug/ml that caused 90% cell die after 7-10d was the best selecting one. After selection, cell was cultured by DMEM containing G418 200ug/ml.