Objective This paper intends to explore the effects of overexpression and low expression of SH2-containing protein tyrosine phosphatase 2(SHP2)on serine-threonine protein kinase(Akt)in liver tissue of carbon tetrachloride(CCl4)induced liver fibrosis rats.Methods One hundred and sixty healthy male SD rats were selected and randomly divided into the control group,the model group,the Ad-GFP group,the Ad-SHP2 group,and the Ad-shRNA/SHP2 group,with 32 rats in each group.A rat liver fibrosis model was constructed by intraperitoneal injection of CCl4.The empty virus Ad-GFP expressing green fluorescent protein(GFP),adenovirus Ad-SHP2 expressing wild-type SHP2 and GFP,and adenovirus Ad-shRNA/SHP2 carrying short hairpin RNA(shRNA)targeting SHP2 expressing GFP were injected into the tail vein of rats.Eight rats were randomly selected from each group at 2nd,4th,6th,and 8th week of modeling,and the liver tissue samples were collected.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression levels of SHP2 and Akt in the liver tissues of rats in each group.Western blotting was used to detect the protein expression levels of SHP2,Akt,and phosphorylated Akt(p-Akt)in the liver tissues of rats in each group.H-E staining was used to observe the pathological changes of liver tissue in each group.Masson trichrome staining was used to observe the collagen deposition in liver tissue in each group.Results SHP2-targeted shRNA and wild-type SHP2 gene are successfully introduced into rats with hepatic fibrosis,and the expression of SHP2 in liver tissues of rats is down-regulated and up-regulated.Compared with the model group and the Ad-GFP group,the degree of liver fibrosis in the Ad-SHP2 group aggravated,while the degree of liver fibrosis in the Ad-shRNA/SHP2 group alleviated.The mRNA and protein expression levels of Akt in the liver tissues of rats in each group are compared,as well as the expression levels of p-Akt protein at the same modeling time point(2nd,4th,6th,and 8th week).Those in the Ad-GFP group,the Ad-SHP2 group,the Ad-shRNA/SHP2 group,and the model group are significantly higher than the control group(P＜0.05),but the expression of Akt at each time point among the Ad-GFP group,the Ad-SHP2 group,the Ad-shRNA/SHP2 group,and the model group have no significant difference(P＞0.05).Compared with the model group and the Ad-GFP group,the expression level of p-Akt protein in the liver tissue of rats in the Ad-shRNA/SHP2 group is significantly reduced(P＜0.05),while that in the Ad-SHP2 group is significantly increased(P＜0.05).There is no statistically significant difference between the model group and the Ad-GFP group(P＞0.05).Conclusion In the course of CCl4 induced liver fibrosis in rats,over expression of SHP2 in liver tissue can increase Akt activity by promoting Akt phosphorylation,while low expression of SHP2 in liver tissue can decline Akt activity by inhibiting Akt phosphorylation.
SH2-containing protein tyrosine phosphatase 2Liver fibrosisProtein kinase B
万方数据
维普
