Effects Of Benefiting Qi, Expelling Blood Stasis, Dissipating Phlegm And Nourishing Yin Method On TGF-β1, TGF-β1R And Liver Tissue Structure Of Rats With Hepatic Fibrosis

Research background:Hepatic fibrosis is a kind of pathological process in which a variety of pathogenic factors induce intrahepatic connective tissue dysplasia, resulting in excessive precipitation of intrahepatic diffuse extracellular matrix(ECM). As a chronic, progressive, diffuse liver lesion, it can develop into cirrhosis if not treated timely or properly. Liver fibrosis is an inevitable phrase from a kind of chronic liver disease developing into cirrhosis, and hepatocellular carcinoma at last. Although gaining its diagnosis name from modern medicine, liver fibrosis, in traditional Chinese medicine, according to its clinical symptoms, has been long before included in the pathological characteristic of hypochondriac pain, jaundice, lumps in the abdomen, tympanites, aggregation-accumulation etc. Its pathogenesis evolution is a dynamic change from exterior excess to interior deficiency, from aggregation to accumulation, from blood to collaterals through qi, whose key mechanism is qi deficiency with blood stasis and phlegm turbidity together. Over years, traditional Chinese medicine(TCM) has achieved good clinical results in the treatment of liver fibrosis through methods of benefiting qi, expelling blood stasis, dissipating phlegm and nourishing Yin respectively or jointly. But, whether the treatment of using the four foresaid therapeutics jointly is better than that of using any of them respectively still remains unproven in present TCM field. So, how to use more scientific means and methods to reveal effects of TCM treatments on liver fibrosis, and simultaneously to more productively guide the clinical treatment of traditional Chinese medicine has become one of important scientific problems today.Objective:To observe effects of the benefiting qi, expelling blood stasis, dissipating phlegm and nourishing yin method on TGF-β1, TGF-β1R and liver tissue structure of rats with hepatic fibrosis induced by subcutaneous injection of CCl4, low protein and high fat diet and alcohol (5%) drinking. To explore the effects of the benefiting qi, expelling blood stasis, dissipating phlegm and nourishing yin method, and the synthetical therapeutics of benefiting qi, expelling blood stasis, dissipating phlegm and nourishing yin, and provide better experimental basis for clinical treatment of liver fibrosis.Methods:1. Establishment of rat models with liver fibrosis and then intervention: SD rats were randomly divided into7groups, including the normal control group, the model group with liver fibrosis, the benefiting qi group, the expelling blood stasis group, the dissipating phlegm group, the nourishing yin group and the benefiting qi&expelling blood stasis&dissipating phlegm with nourishing yin group. Except the normal control group, the rest groups were all made rat model groups with liver fibrosis by subcutaneous injection of CCl4, low protein+high fat diet and alcohol (5%) drinking Then all7groups were treated accordingly and separately with normal saline, the benefiting qi decoction, resolving blood stasis decoction, the dissipating phlegm decoction, the nourishing yin decoction, and the benefiting qi& expelling blood stasis&dissipating phlegm with nourishing yin decoction. On the14th weekend, specimens were correspondingly made based on testing requirements through methods of cardiac perfusion, taking livers directly from rats etc.2. Pathological change and ultrastructure in hepatic tissues were observated by light microscopy,Masson collagen staining method and electron microscopy techniques.3. TGF-β1and TGF-β1R mRNA and protein expression in liver tissues of each group were observated by RT-PCR and Western Blot methods.Results:1. Compared with the normal control group, the degrees of liver fibrosis in rats from model groups were significantly higher and the TGF-β1and TGF-β1R mRNA and protein expression significantly increased (P<0.01)2.. By the comparison, method of benefiting qi, expelling blood stasis, dissipating phlegm and nourishing yin and benefiting qi&expelling blood stasis&dissipating phlegm with nourishing yin method all can reduce degree of the liver fibrosis and mRNA and protein expression of TGF-β1and TGF-β1R (P<0.01) in hepatic tissues, while the benefiting qi&expelling blood stasis&dissipating phlegm with nourishing yin method is best (P<0.05)Conclusion:Method of benefiting qi, expelling blood stasis, dissipating phlegm and nourishing yin and benefiting qi&expelling blood stasis&dissipating phlegm with nourishing yin method all can reduce degree of the liver fibrosis in liver tissues of rats with hepatic fibrosis. Its mechanisms may relate to decrease mRNA and protein expression of TGF-β1and TGF-β1R (P<0.01) in hepatic tissues.