Objective To investigate the effect of overexpression of Src homology 2 domain-containing protein tyrosine phosphatase 2(SHP2)on apoptosis in human hepatic stellate cells(LX-2).Methods LX-2 cells were transfected with adenoviruses expressing green fluorescent protein(GFP)alone(Ad-GFP),wild-type SHP2 and GFP(Ad-SHP2).The experiment was divided into three groups:Control group,in which LX-2 cells were transfected with DMEM instead of adenovirus;Ad-GFP group,transfected with empty adenovirus Ad-GFP;Ad-SHP2 group,transfected with recombinant adenovirus Ad-SHP2.Western blotting was used to detect the expression of SHP2,Bax,and Bcl-2 proteins in the three groups of LX-2 cells.Real-time quantitative polymerase chain reaction(qRT-PCR)was performed to measure SHP2 mRNA expression in LX-2 cells.Apoptosis of LX-2 cells in the three groups was detected by TUNEL assay and annexin V/propidium iodide double staining flow cytometry.Results The relative expression levels of SHP2 protein and mRNA in LX-2 cells in the Ad-SHP2 group were higher than those in the Control group and Ad-GFP group(P＜0.05).There was no significant difference in SHP2 expression between the Control group and Ad-GFP group(P＞0.05).TUNEL assay and annexin V/propidium iodide double staining flow cytometry showed that the apoptosis rate of LX-2 cells in the Ad-SHP2 group(1.52±0.26)%and(2.44±0.93)%was lower compared to the Control group(3.08±0.73)%and(9.44±0.80)%and the Ad-GFP group(3.25±0.85)%and(8.89±1.98)%(P＜0.05).There was no significant difference in apoptosis between the Control group and Ad-GFP group(P＞0.05).The relative expression levels of Bax and Bcl-2 proteins in LX-2 cells in the three groups were significantly different(P＜0.05).The relative expression level of Bax protein in the Ad-SHP2 group(1.55±0.21)was lower than that in the Control group(1.99±0.15)and the Ad-GFP group(2.00±0.16)(P＜0.05),while the relative expression level of Bcl-2 protein(2.13±0.25)was higher than that in the Control group(1.71±0.15)and the Ad-GFP group(1.52±0.14)(P＜0.05).There was no significant difference in the expression of Bax and Bcl-2 proteins between the Control group and Ad-GFP group(P＞0.05).Conclusion Overexpression of SHP2 inhibits apoptosis of human hepatic stellate cells LX-2 in vitro by increasing the Bcl-2/Bax ratio.
维普
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