Effects Of Hydrogen Sulfide On Hepatic Fibrosis In Rats And Its Relationsh IP With The P38MAPK Signal Pathway

Objective:To investigate the effects of sodium hydrosulfide (NaHS) as the donor of hydrogen sulfide (H2S) andSB230580as the inhibitor of P38MAPK signal pathway respectively on liver tissue morphology andcollagen deposition in hepatic fibrosis rat models. To detect the relationship between H2S and P38MAPKsignal pathway for better understanding the mechanism of hydrogen sulfide on hepatic fibrosis rats.Method:The hepatic fibrosis rat models were established by hypodermic injection of carbon tetrachloride mixedwith cottonseed oil at the concentration of40%, feeding high-fat, high-cholesterol diet and drinking ethanol.The rats were randomly divided into five groups after succeed establishing models: hepatic fibrosis group(group HF), DMSO group (group D), NaHS group (group S),SB203580group (group SB), andSB203580+NaHS group (group SB+S). The rats in group HF, group D, group SB and group S wereintraperitoneally infused with physiologic saline,2‰DMSO solution, SB203580solution (0.3mg/kg·d),and NaHS solution (56μmol/kg·d) separately for4times. The rats in group SB+S were intraperitoneallyinfused with SB203580solution (0.3mg/kg·d) and NaHS solution (56μmol/kg·d) simultaneously for4times.All rat livers were collected after all above treatments were finished. Hepatic fibrosis pathology stages weredetermined by HE staining. The depositions of collagen fiber were observed by Masson staining. Theexpressions of type I and III collagen were tested by RT-PCR and immunohistochemisty.The expressions of P38MAPK and caspase-3were tested by western blot.Result:(1)HE staining confirmed hepatic cells degeneration and necrosis, hyperplasia of fibrous connective tissue,and the formation of false lobular and infiltration of inflammation cells.(2)HE staining was used todetermine hepatic fibrosis stages. Compares with group N, the stage of hepatic fibrosis raised apparently ingroup HF and group D. Compared with group HF and group D, the stage of hepatic fibrosis in group S,group SB and group SB+S were decreased.There was no obvious difference among group SB and group S.But compared with group S and group SB, the stage of hepatic fibrosis in group SB+S decreased.(3)Masson staining was used to calculate the score according to fibrosis semi-quantitative scoring system inliver to observe the deposition of collagen fiber. The fibrosis semi-quantitative score of group HF andgroup D were remarkable higher than group N. The fibrosis semi-quantitative score of group S,group SBand group SB+S were lower than group HF and group D. The fibrosis semi-quantitative score of groupSB+S was lower than group S and group SB, but there was no obvious difference between group SB andgroup S.(4)Immunohistochemical staining and RT-PCR were used to detected type I and III collagenprotein expression and mRNA expression. Type I and III collagen protein expression and mRNAexpression were increased significantly in group HF and group D than those of group N. Compared withgroup HF and group D, Type I and III collagen protein expression and mRNA expression were decreasedin group S,group SB and group SB+S. Type I and III collagen protein expression and mRNA expression ingroup SB+S was less than group S and group SB, but there was no obvious difference between group SBand group S.(5)Western blot results showed that there was no obvious difference among group N, groupHF, group D and group S about the expression of P38MAPK, in group HF expressed more than group SB,but in group SB expressed less than group SB+S. In group HF, caspase-3expressed more than group N,group S, group SB and group SB+S. In group SB+S, caspase-3expressed more than group S and group SB,but capase-3expressed in group S less than group SB.Conclusion:(1)Carbon tetrachloride combined with ethanol, high-fat and high-cholesterol diet could establish hepaticfibrosis rat models successfully.(2) P38MAPK signal pathway was closely related to the development ofhepatic fibrosis and its inhibitor SB203580could significantly improve hepatic fibrosis.(3)These protectiveeffects of hydrogen sulfide postconditioning on hepatic fibrosis might be related to P38MAPK signalpathway.