Glucagon like peptide-1 mediated PI3K/Akt pathway antagonizes advanced oxidized protein products induced apoptosis of HTR-8/SVneo cells / 中国医师杂志

ABSTRACT

Objective:To explore the protective mechanism of phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in glucagon like peptide-1 (GLP-1) antagonizing the apoptosis of gestational trophoblasts (HTR-8/SVneo) induced by advanced oxidized protein products (AOPP).Methods:Pregnant trophoblast HTR-8/SVneo were cultured in vitro. The cells were divided into control group, AOPP group, GLP-1 group, AOPP + GLP-1 group and AOPP + GLP-1 + LY294002 group. The control group was cultured in 1640 medium; AOPP group was stimulated with 200 μg/ml AOPP; GLP-1 group was stimulated with 50-100 nmol/L GLP-1 for 1 h; AOPP + GLP-1 group was stimulated with 200 μg/ml AOPP for 48 hours, and then GLP-1 (50-100 nmol/L) was added for 1 hour; In AOPP + GLP-1 + LY294002 group, PI3K inhibitor LY294002 was added on the basis of the intervention of AOPP + GLP-1 group. The expression of PI3K/Akt pathway related protein p-Akt was detected by Western blot. Cell viability was detected by cell counting kit (CCK-8). Enzyme linked immunosorbent assay (ELISA) was used to detect the contents of apoptosis promoter protease caspase-9 and caspase-3, and the contents of apoptosis related proteins Bcl-2, Bax and Cyto-c. Results:After AOPP stimulation, the expression of p-Akt in AOPP group was lower than that in control group ( P<0.05); After 50 and 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After 24 and 48 hours of 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After AOPP stimulation, the cell viability of AOPP group was lower than that of control group ( P<0.05); After GLP-1 intervention, the cell viability of AOPP + GLP-1 group was significantly higher than that of AOPP group ( P<0.05). After adding PI3K inhibitor LY294002, the cell viability of AOPP + GLP-1 + LY294002 group was significantly lower than that of AOPP + GLP-1 group ( P<0.05). The results of ELISA showed that the contents of apoptosis promoter protein caspase-3, caspase-9, apoptosis related protein Bax and Cyto-c in AOPP group were higher than those in control group (all P<0.05), and the content of anti-apoptosis protein Bcl-2 was lower than that in control group ( P<0.05); After GLP-1 intervention, the contents of caspase-3, caspase-9, Bax and Cyto-c in AOPP + GLP-1 group were significantly lower than those in AOPP group ( P<0.05), and the content of anti-apoptosis protein Bcl-2 was higher than that in AOPP group ( P<0.05). After treatment with PI3K inhibitor LY294002, the contents of Bcl-2 in AOPP + GLP-1 + LY294002 group were lower than those in AOPP + GLP-1 group, and the contents of Bax and Cyto-c were higher than those in AOPP + GLP-1 group (all P<0.05). Conclusions:GLP-1 may mediate PI3K / Akt pathway to antagonize the apoptosis of HTR-8/SVneo induced by AOPP.