Effect of CircACAP2 Targeting miR-29a-3p/CCNT2 Axis on Hypoxia/Reoxygenation Induced Cardiomyocyte Injury
Objective:To investigate the effect of CircACAP2 targeting miR-29a-3p/cyclin T2(CCNT2)axis on hypoxia/reoxygenation(H/R)induced cardiomyocyte injury.Methods:H9c2 cells were divided into Control group(normal culture),si-NC group(transfected with si-NC),Model group(established H/R model),si-CircACAP2 group(transfected with si-CircACAP2),si-CircACAP2+inhibitor-NC group(si-CircACAP2 and inhibitor-NC co-transfection),si-CircACAP2+miR-29a-3p inhibitor group(si-CircACAP2 and miR-29a-3p inhibitor co-transfection).The expression levels of CircACAP2 and miR-29a-3p,cell viability and apoptosis of H9c2 cells were detected.H9c2 autophagy,lactate dehydrogenase(LDH),malondialdehyde(MDA),superoxide dismutase(SOD),reactive oxygen species(ROS)levels and expression of proliferating cell nuclear antigen(PCNA),B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),Beclin-1,P62 CCNT2 were dected.The targeting relationship of CircACAP2 to miR-29a-3p and the targeting relationship of miR-29a-3p to CCNT2 were detected.Results:Compared with the Control group,the expression of CircACAP2,apoptosis rate,number of autophagy vesicles,LDH release rate,MDA,ROS levels and Bax,Beclin-1,CCNT2 protein increased in H9c2 cells in Model group,the expression of miR-29a-3p,OD490 value,SOD activity,PCNA,Bcl-2 and P62 protein expression decreased(P<0.05).Compared with Model group,CircACAP2 expression,apoptosis rate,number of autophagy vesicles,LDH release rate,MDA,ROS levels,Bax,Beclin-1 and CCNT2 protein expression in H9c2 cells of si-CircACAP2 group decreased,miR-29a-3p expression,OD490 value,SOD activity,PCNA,Bcl-2,P62 protein expression increased(P<0.05).Compared with si-CircACAP2 group,there was no statistically significant difference in CircACAP2 expression in H9c2 cells of si-CircACAP2+miR-29a-3p inhibitor group(P>0.05),apoptosis rate,number of autophagy vesicles,LDH release rate,MDA,ROS levels and Bax,Beclin-1,CCNT2 protein expression increased,and miR-29a-3p expression,OD490 value,SOD activity,PCNA,Bcl-2,P62 protein expression decreased(P<0.05).CircACAP2 targeted the expression of miR-29a-3p,and miR-29a-3p negatively regulated the expression of CCNT2.Conclusion:Knockdown of CircACAP2 may down-regulate CCNT2 expression by targeting miR-29a-3p,thereby inhibiting H/R-induced apoptosis,autophagy and oxidative stress of H9c2 cells,promoting cell proliferation and playing a protective role.
anoxia/reoxygenationCircACAP2miR-29a-3p/cyclin T2 axismyocardial cellsexperimental study
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