| Background and Objective:One of the most important issues affecting the global public health belongs to cirrhosis,its etiology is various,among them,died of liver cirrhosis,liver failure and hepatocellular carcinoma caused by HBV infection about 1million people worldwide each year [1],the clinical focus on etiology and symptomatic treatment of liver cirrhosis,but liver cirrhosis is irreversible lesions,therefore,it is very important for the early lesions of liver cirrhosis the stage of liver fibrosis intervention.The liver,s immune responses to various external stimuli is a key factor in disease progression and clinical outcome of liver,then based on the original treatment combined with new immunotherapeutic strategies may be a useful method for reversion of liver fibrosis.Studies have shown that the new immune factor IL-17 may be involved in the occurrence of multiple organ fibrosis,and relevant experiments have confirmed that rapamycin can regulate the balance of Th17/Treg cells to improve CCl4 induced liver fibrosis in rats.The research team found that pre experiment,active vitamin D3 as an immunomodulator,via inhibiting HSC proliferation and apoptosis play a role of anti fibrosis,but the mechanism is not clear,not widely used in clinical application.In this study,we investigated the mechanism of 1,25-(OH)2 D3(1,25-dihydroxyvitamin D3)in order to prevent the occurrence and development of hepatic fibrosis.Through observing the influence on gene and protein levels expression of IL-17 and its related factors of ROR?t and MIP3? in the progression of hepatic fibrosis in rats,this experimentanalysised the possible mechanism of 1,25(OH)2D3 inhibiting rat liver fibrosis development.Methods:The model of liver fibrosis rats(Sprague-Dawley)were established by the classic method of intraperitoneal injection of CCL4.In the treatment group were alsoreceived 1,25(OH)2D3peanut oil solution throughgavage for 8weeks.By using liquid chromatography tandem mass spectrometry detection of serum 25(OH)D3 concentrationdischarge immunoassay of serum liver fibrosis four,enzyme linked immunosorbent assay(ELISA)dynamic analysis of serum interleukin 17 concentration,the difference of serum liver fibrosis four and IL-17 expression was observed between normal group and model group,drug solvent group and treatment group.By HE and Masson staining of liver tissue of rats in each group to divide the stage of pathological,immunohistochemical method to detect the liver tissue IL-17 and MIP3? protein expression,Western blot assay and real-time quantitative immunofluorescence(RT-PCR)method to determine the expressionlevel of IL-17,ROR?t,MIP3? protein and mRNA in liver tissue,hepatic fibrosis and the expression of hepatic IL-17,ROR?t,MIP3? protein and mRNAwas observed in normal group and model group,drug solvent group and treatment group.Results:1.Pathological examination showed that the model was established successfully,the treatment group rats serum 25(OH)D3 concentration was significantly higher than the drug solvent group(P=0.000 < 0.05);model group and drugsolvent group serum 25(OH)D3 levels were significantly lower than the normal(P < 0.05),the differences were significantstatistically.2.At 8 weeks,the expression of the model group and the drug solvent group serum HA,LN,P?NP,CIV and IL-17 were significantlyhigher than the normal group,and the drug solvent group wassignificantly higher than the treatment group(P < 0.05),the differenceswere significantstatistically.In the process of modeling,the serum IL-17 level was decreased first but increased with the time after.3.Through the HE and Masson staining of liver tissue of rats in each group,we found that compared with normal group(S0),model group(S4)and drug solvent group(S4)in rats with liver fibrosis significantly,while the degree of treatment group(S2)hepatic fibrosis was more than that of the model group and drug solvent group.4.The model group and the drug solvent group was significantly higher than that of IL-17,ROR?t,MIP3 ? protein expression in the treatment group in the liver of rats(P < 0.05),the difference were significantstatistically,at the same time,in the modeling process,the expression of IL-17 protein with time gradually increased in liver tissue showed a time dependent.5.The detection of liver tissue of rats by Rt-PCR method,found that the rats in the normal group,the liver IL-17 ROR?t and MIP3?mRNA expression is minimal,and the model group and drug solvent group liver IL-17,ROR?t and MIP3? mRNA expression is significantly higher than the treatment group(P < 0.05),the difference has statistical significance.6.Analysis of the correlation of the expression of IL-17 in rat serum and liver tissue at different time points(4W,6W,8W),found the expression of IL-17 in serum and liver tissue in model group,drug solvent group and the treatment group was negatively correlated over time(r1=-0.979,r2=-0.611,r3=,-0.878).Conclusion:1.the expression of IL-17 in serum and liver of rats with hepatic fibrosis was significantly higher than that in normal rats,suggesting that IL-17 may be involved in the development of hepatic fibrosis;2.1,25(OH)2D3 with on hepatic fibrosis in rats was significantly inhibited,further confirmed that 1,25-dihydroxyvitamin D3 inhibits hepatic fibrosis;3.1,25(OH)2D3 decreased IL-17 and its related factors ROR?t,MIP3?expression of protein and mRNA significantly,and played the role of anti fibrosis,suggesting that 1,25(OH)2D3 may regulateactivationandproliferation of rat HSC through immune micro environment mediated by IL-17. |
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