The Impacts of Stachydrine on H/R Induced Apoptosis of Myocardial Cellsby Regulating the EPAC1/Rap1 Signaling Pathway

The aim of this study was to investigate the impacts of STA (stachydrine) on H/R (hypoxia/reoxygenation) induced apoptosis of myocardial cells by regulating the EPAC1 (exchange protein 1 directly activated by cAMP)/Rap1 (Ras associated protein 1) signaling pathway. H9C2 cells were grouped into control group,model group (H/R), H/R+STA (5 μmol/L STA) group, H/R+8-CPT (EPCA1 agonist 8-CPT, 30 μmol/L) group,H/R+STA+8-CPT group (5 μmol/L STA+30 μmol/L 8-CPT), H/R+ESI-09 (EPCA1 inhibitor ESI-09, 1 μmol/L)group, and H/R+STA+ESI-09 (5 μmol/L STA+1 μmol/L ESI-09) group; CCK-8 method was applied to detect theproliferation of H9C2 cells; DCFH-DA method was applied to detect the level of ROS (reactive oxygen species) inH9C2 cells; flow cytometry was applied to detect H9C2 cell apoptosis; qRT-PCR (real-time fluorescence quantitative PCR) was applied to detect the mRNA expression levels of EPAC1 and Rap1 in myocardial cells; Western blotwas applied to detect the expression levels of EPAC1, Rap1, Bax (Bcl-2 associated X pretein), and Bcl-2 (B celllymphocyte tumor 2) proteins in myocardial cells. Compared with the control group, the level of ROS, apoptosisrate, the protein expression of Bax, as well as the mRNA and protein expression of EPAC1 and Rap1 in H9C2 cellsin the H/R group were obviously increased, while the cell proliferation activity and the protein expression of Bcl2 were obviously reduced (P<0.05); compared with the H/R group, the level of ROS, apoptosis rate, the proteinexpression of Bax, as well as the mRNA and protein expression of EPAC1 and Rap1 in H9C2 cells in the H/R+STAgroup and H/R+ESI-09 group were obviously reduced, while the cell proliferation activity and the protein expression of Bcl-2 were obviously increased (P<0.05); the indicators in the H/R+8-CPT group showed opposite trends(P<0.05); compared with the H/R+STA group, the level of ROS, apoptosis rate, the protein expression of Bax, aswell as the mRNA and protein expression of EPAC1 and Rap1 in H9C2 cells in the H/R+STA+8-CPT group wereobviously increased, the cell proliferation activity and the protein expression of Bcl-2 were obviously reduced(P<0.05), the indicators in the H/R+STA+ESI-09 group showed opposite trends (P<0.05). In conclusion, STA mayimprove H/R-induced cardiomyocardial cells apoptosis by inhibiting the EPAC1/Rap1 signaling pathway.

CSTR: 32200.14.cjcb.2023.11.0003