The Effect Of Yupingfeng Pachy-polysaccharide On TGF-β1-induced Rat Hepatic Stellate Cell And The Relationship With Smads Signaling Pathway

Liver fibrosis is the development of most chronic liver diseases,at the same time, is also common to the pathological process of cirrhosis, chronic liver injury is healing response for a reversible process. At present, the incidence of various liver diseases is rising, they are accompanied by hepatic fibrosis ( hepatic fibrosis, HF), eventually develop into cirrhosis or even cause liver failure and then lead to death, that is the major diseases harm to the health of people worldwide. And a long time, there is no clinical curative effect of drugs to prevent its occurrence and development. Thus, people are paying more and more attention to liver fibrosis researchers.Hepatic fibrosis is the excessive deposition of fibrous connective tissue of liver, and is the result of the extracellular matrix synthesis and degradation balance, moreover is the pathological changes and the common necessary way of the development of chronic liver disease to cirrhosis. Hepatic fibrosis is due to various factors cause liver cell damage, necrosis, apoptosis and liver inflammation, activation of Kupffer cells to secrete cytokines, and then act on hepatic stellate cells together. The feature of HSC activation can reflect byα-SMA and ECM increased generation. Transforming growth factor-beta activated hepatic stellate cell, with activation of HSC, the promotion of collagen gene expression in the liver. TGF-β1 depends on the receptor of downstream and intracellular signal that is highly conserved family of Smad proteins mainly, mediates the signal transduction, and plays a biological effect. Recently someone discover a growth factor-CTGF which is the downstream effect media of TGF-β1. It can promote the proliferation of fibroblasts, and participate in the generation and accumulation of ECM. CTGF itself does not inhibit the anti-inflammatory and immune, and is not a substitute for TGF-β1. It is only mediated biological functions. Therefore, there may be a new target for liver fibrosis by influencing or regulating the expression of CTGF.Yu Ping Feng San composed of Astragalus, Atractylodes, and wind, are widely used to prevent cold cold, upper respiratory tract infections, allergic diseases, allergic diseases, bronchial asthma and other diseases, not only show a significant effect but also play a positive role in the fight against SARS. Early results show that, YPF-P has a protective effect on acute experimental liver injury in mice, and there are good preventive effects on carbon tetrachloride (CCl4) induced liver fibrosis.Nowadays, it is recognized that liver fibrosis is reversible, but there are still many issues to be resolved, liver fibrosis is interwoven by a variety of complex factors. At present, To begin with the development of hepatic fibrosis from different sectors, many anti-fibrosis drugs are developed. But still no determined, effects of drugs, the treatment of liver fibrosis is provided a new progress by all the clarification of the pathogenesis of hepatic fibrosis.To further clarify the mechanism of liver fibrosis Yupingfeng polysaccharides affct on, the present study adopts the experiment that YPF-P affect on the cells cultured in vitro directly. In order to discuss the effct of Yupingfeng polysaccharide on TGF-β1-induced hepatic stellate cell proliferation and the Smads protein And its mediated signaling pathway in hepatic fibrosis, the role in the development of the preliminary,the research can be devided into two parts:1. Effct of Yupingfeng polysaccharides on the proliferation of TGF-β1-induced rat hepatic stellate cell andα-SMA and the expression of t collagenⅠ, collagenⅢmRNAHepatic stellate cell strains HSC-T6 were divided into 7 groups: normal group, model group, YPF-P (12.5mg / L, 25 mg / L, 50mg / L, 100mg / L, 200mg / L), then used the MTT assay to detect the condition of HSC cell proliferation to screen the most appropriate dose and time. Semi-quantitative RT-PCR was used to detect theα-SMA and collagenⅠ, collagenⅢmRNA of HSC. MTT results showed that, TGF-β1 could promote the proliferation of HSC. YPF-P25, 50,100 mg/L could well inhibit TGF-β1-induced the proliferation of HSC. And the most obvious effect was the 48h. RT-PCR results showed that, Each dose group of YPF-P could significantly inhibit the expression ofα-SMA and collagenⅠ, collagenⅢmRNA. To the point, YPF-P could inhibit the activation of HSC proliferation and collagen secretion.2. Effect of Yupingfeng polysaccharide on the expression of CTGF and Smad signal transduction pathway in TGF-β1-induced rat hepatic stellate cellsHepatic stellate cell strains HSC-T6 was dealed with the YPF-P (25, 50, 100 mg/L) for 48 hours, while setting blank control and TGF-β1-induced control, semi-quantitative RT-PCR detected the expression of Smad2, Smad3, Smad7 And CTGFmRNA in HSC. The results showed that, Each dose group of YPF-P could lower the expression of Smad2, Smad3, CTGFmRNA, increased the expression of Smad7mRNA. YPF-P could inhibit the activation of HSC proliferation and collagen secretion, which might be involve with blocking the Smads signaling pathway in HSC.Based on the above, HSC-T6 were dealed with YPF-P (50mg/L), blocking agent SIS3(10μmol/ml) and YPF-P (50mg/L)+ blocking agent SIS3(10μmol/ml) for 48 hours, and meanwhile compared with blank and TGF-β1-induced group. The expression of CTGF protein in HSC was observed by Western blot. The strongest expression of protein CTGF was the model group, and by turns were normal group, medication group, blocking agent group and blocking agent-medication group. YPF-P could inhibit the activation of HSC proliferation and collagen secretion. The YPF-P's mechanism might be associated with blocking the Smads signaling pathway and then reduced the expression of CTGF.