| Objective:To investigate the effects of sodium hydrosulfide (NaHS) as the donor of hydrogen sulfide (H2S) andLY294002as the inhibitor of PI3K/Akt signal pathway respectively on liver tissue morphology andcollagen deposition in hepatic fibrosis rat models. To detect the relationship between H2S and PI3K/Aktsignal 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 six weeks: hepatic fibrosis group (group HF), DMSOgroup (group D), LY294002group (group L), NaHS group (group S), and LY294002+NaHS group (groupLS). The rats in group HF, group D, group LY and group S were intraperitoneally infused with physiologicsaline,2‰DMSO solution, LY294002solution (0.3mg/kg·d), and NaHS solution (56μmol/kg·d) separatelyfor12times. The rats in group LS were intraperitoneally infused with LY294002solution (0.3mg/kg·d) andNaHS solution (56μmol/kg·d) simultaneously for12times. All rat livers were collected after all above trea-tments were finished. Hepatic fibrosis pathology stages were determined by H&E staining. The depositionsof collagen fiber were observed by Masson staining. The expressions of type I and III collagen were testedby RT-PCR and immunohistochemisty. The expressions of PI3K and p-Akt were tested by western blot.Result:(1)H&E staining confirmed hepatic cells degeneration and necrosis, hyperplasia of fibrous connectivetissue, and the formation of false lobular and infiltration of inflammation cells.(2)H&E staining was usedto determine hepatic fibrosis stages. Compares with group N, the stage of hepatic fibrosis raised apparentlyin group HF and group D. Compared with group HF and group HF and group D, the stage of hepaticfibrosis in group S and group LY were decreased. But there was no obvious difference among group LY,group S and group LS.(3)Masson staining was used to calculate the score according to fibrosissemi-quantitative scoring system in liver to observe the deposition of collagen fiber. The fibrosissemi-quantitative score of group HF and group D were remarkable higher than group N. The fibrosissemi-quantitative score of group S and group LY were lower than group HF and group D. The fibrosissemi-quantitative score of group LS was lower than group S, but higher than group LY.(4)Immunohistochemical staining and RT-PCR were used to detected type I and III collagen proteinexpression and mRNA expression. Type I and III collagen protein expression and mRNA expression wereincreased significantly in group HF and group D than those of group N. Compared with group HF andgroup D, Type I and III collagen protein expression and mRNA expression were decreased in group S andgroup LY. Type I and III collagen protein expression and mRNA expression in group LS was less thangroup S, but more than group LY.(5)Western blot results showed that PI3K and p-Akt in group HF andgroup D expressed more than group N, but these two proteins in group LY expressed less than group D.These proteins had no obvious difference between group S and group HF. In group LS, PI3K and p-Aktexpressed more than group LY, but less than group S.Conclusion:(1)Carbon tetrachloride combined with ethanol, high-fat and high-cholesterol diet could establish hepaticfibrosis rat models successfully.(2)Hydrogen sulfide delayed the progress of hepatic fibrosis and hadprotective effects on the hepatic fibrosis by inhibiting morphology damage and decreasing type I and IIIcollagen expression.(3)PI3K/Akt signal pathway was closely related to the development of hepatic fibrosisand its inhibitor LY294002could significantly improve hepatic fibrosis.(4)These protective effects ofhydrogen sulfide postconditioning on hepatic fibrosis might be related to PI3K/Akt signal pathway. |
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