| Background:Cirrhosis is a worldwide disease which seriously threatens people’s health. Almost all chronic liver disease associated with liver fibrosis. Currently consider that liver fibrosis is a kind of disease.As a kind of disease (K74.001) in International Classification of Diseases (ICD-10). A growing body of research suggests that reasonable treatment can restore the fibrosis of the liver, and even to cure. Current research for the prevention and treatment of liver fibrosis is still in the basic level, and lack of specific effective drug intervention of liver fibrosis in clinical. Rhodiola genus crassulaceae plants, and Salidroside is one of the effective components. Studies have shown that it has antioxidant properties. In recent years,studies have shown that it has better anti-fibrosis action,but the mechanism is still unclear. Therefore.we establish a mouse liver fibrosis model induced by CCL4and by blocking the ROS pathway, to observe the effects of salidroside on liver fibrosis and the related signal pathways in mice, then reveal the possible mechanism of action. For the clinical application of salidroside provide some experimental evidence.Part one:The effects of Sal on CCL4induced liver fibrosis in miceObjective:To observe the effect of salidroside on liver fibrosis, and investigate the influence of ROS-related pathways involve TGF-β1, NF-kB, MMPs/TIMPs.Method:the liver-fibrosis model was produced by intraperitoneal injection of CCL4in mice. Respectively given Sal and N-acetylcysteine (NAC) processing. After8weeks, execution the mice and then collect its serum,liver, at the same time detection of MDA, SOD, GSH. Liver were observed in HE and masson staining. Extraction of liver tssue’s RNA examination the mRNA expression level of MMP-9/TIMP-1,TGF-β1. Extract the HSCs primary cell culture, detected the MMP-9/TIMP-1,TGF-β1mRNA expression level in HSCs. Western-blot detection the protein expression level of NF-κB P65.Result:Pathological and immunohistochemical staining observed that the degree of liver fibrosis was significantly reduced in Sal and NAC groups. In serum,the ALT, AST, MDA, TGF-β1levels decreased (P<0.01)and the SOD levels increased (P<0.01)in Sal、NAC treatment groups. Compare with CCL4model groups, GSH, GSH-Px, SOD levels increased(P<0.01), MDA decreased in liver tissue(P<0.01).RNA was extracted from liver tissue and HSCs primary cell detection that Sal, NAC treatment groups compared with CCL4model groups of MMP-9mRNA level without statistical significance and TIMP-1, TGF-β1mRNA level decreased(P<0.01). Western-blot detected the NF-κB P65protein levels, Sal, NAC treatment group compared with CCL4model group decreased(P<0.01).Conclusion:Sal has obvious anti liver fibrosis effect,its mechanism of action involves the ROS related TGF-beta1, MPP/TIMP, the NF-κB pathwayPart two:Sal inhibits LPS induced proliferation in HSC-T6by NO pathwayObjective:To investigate the effect of Salidroside on proliferation of hepatic stellate Cells(HSCs) and estimate the function of the NO during the procedure.Methods:Choosing the lipopolysaccharide(LPS)used to activate HSC-T6.Rat HSC-T6activated by LPS were incubated with various concentration of Sal. Then conduct the following experiment after twenty-four hours:MTT colorimetric assay was used to detect the Proliferation of HSC-T6;The intracellular NO was detected using DAF-FM DA.Western blot were used to investigate the expression of iNOS protein.Results:Compared with the control group, the ratio of cell proliferation increased(P<0.01) after LPS-induced HSC-T6for24h. Compared with the LPS group, the ratio of cell proliferation decreased(P<0.01) after Sal treatment with LPS-induced HSC-T6for24h,and in a dose-dependent manner. In group treated with Sal, the NO concentration and expression level of iNOS in HSC-T6increased (P<0.01), and in a dose-dependent manner(P<0.01).Conclusion:Sal could inhibit the proliferation of HSC-T6.which could be mediated by NO partially at least. |
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