Mechanisms For The Antifibrotic Action Of Mushu Capsule On Rats And Its Curative Effect On Patients With Liver Cirrhosis

[background and objective]hepatic fibrosis is a pathological change which associated with the unbalance of extracellular matrix proliferation and degradation and the deposition of connective tissues when parenchymal hepatic cell necrosis and inflammation happened. In recent years , the theory which hepatic fibrosis would be reversed was raised, so the study of hepatic fibrosis became a topic in the domain in world. Traditional Chinese medicine have priority of treatment on hepatic fibrosis in many ways and many levels.We have proved that mushu capsule can improve liver function of hepatic fibrosis rats and can depress the the serum markers of hepatic fibrosis ,degrade the pathological changes , delay the development of liver fibrosis at a preventative dosage. We have further proved that the protective mechanism maybe related with the anti-oxidation and down-regulation of TIMP-1. Based on the above work, we need to carry out more academic analysis and explanation about the attracting anti-fibrosis mechanism of mushu capsule using biomolecular methods. In this study, we firstly investigate the mechanism of reversing hepatic fibrosis in rats by mushu capsule, and then observe its therapeutic effects in patients with liver cirrhosis. [Materials and Methods]Part One: Wistar rats were randomly divided into 4 groups: normal control group, model group, mushu capsule group and ruanganpian group. Mushu capsule group was subdivided into prevention group and treatment. Hepatic fibrosis modelwas induced by subcutaneous injection of CCL4 (40% density, 0.03ml/kg body weight) twice a week for eight weeks. Rats in prevention group were given drugs at first injection of CCL4 . Rats in treatment group were given drugs after injection of CCL4. At the end of the experiment, all the rats were sacrificed. The rat blood was drawn from the heart and the livers were removed. Determination of serum interluekine(IL) 2,6,8 and tumor necrosis factor-a(TNF-a) were performed and the expression of TNF-a was analyzed by RT-PCR, at the same time, tissue sections were stained respectively through HE and Masson method. Macro and micro features changes of the rat's liver were observed. The expression of CD4 and CD8 was detected by flow cytometry.Part Two: The Mechanism of Mushu capsule reversing liver fibrosis in rats. RT-PCR was used to detect the expression of TGFB1 > a-SMA, Col-1 , Col-III in fibrotic liver.Part Three: The information of each patients' complains and symptoms were collected. In order to research the treatment effect of mushu capsule on patients with liver cirrhosis, serum liver function was tested on the lth, 2th and 3th month after the first mushu capsule treatment respectively. [Results]Part One: 40% CCL4 injection could induce rat hepatic fibrosis. All the rats had hepatic fibrosis in the end.Part Two: The elevated serum IL-6, IL-8 and TNF-a were markedly decreased, and the decreased IL-2 was improved after the treatment of mushu capsule and ruanganpian. There was no difference between mushu capsule and ruanganpian in altering IL-2 and IL-6, while there was significant difference in terms of IL-8. The elevated serum TNF-a was decreased more noticeable by mushu capsule preventing group than ruanganpian, the elevated expression of it was markedly decreased by mushu capsule and ruanganpian. The decreased serum CD4 and CD4/CD8 were markedly raised in mushu capsule preventing groups and ruanganpian preventing group. The pathology change of collagenic fibers in liverwas alleviated in mushu capsule prevention groups. The severity of fibrosis was not significantly decreased in mushu capsule treatment groups compared with CCL4 group.Part Three: The elevated expression of TGF81 > a-SMA, Col-1 > Col-III were decreased by mushu capsule and ruanganpian. There was significant difference between mushu capsule treatment group and ruanganpian group.Part Four: Symptoms of flatulence, poor appetite and fatigue in patients with liver cirrhosis were alleviated after mushu capsule treatment. The decreased prothrombin time and hemoglobin were improved and the decreased serum albumin showed an increasing tendency. [Conclusion]All of our results revealed that:1.Preventing treatment of experimental hepatic fibrosis rats with mushu capsule could improve the histopathologic changes and repress the deposition of ECM in liver tissue and then prevent the rats from developing hepatic fibrosis. The mechanisms of mushu capsule in protecting the Wister rats from CCLrinduced hepatic fibrosis might be: ?influencing the function of immune system, ?inhibiting the activation of TGF-81 and a-SMA and then causing the reduction of Col-1 and Col-III .2.Mushu capsule could improve the symptoms of patients with liver cirrhosis and liver synthesizing function.3.Musu capsule have potential value for clinical use.