Effect of circular RNA circHIPK3 on clozapine induced injury of H9C2 myocardial cells and its related mechanisms
Objective To investigate the effect of circular RNA(circRNA)circHIPK3 on clozapine induced injury of H9C2 my-ocardial cells and its related mechanisms.Methods CircHIPK3 was overexpressed or silenced in H9C2 cells by cell transfection and H9C2 cells were treated with clozapine.Real-time quantitative PCR(qRT-PCR)was used to detect the expression of circHIPK3.Methyl thiazolyl tetrazolium assay(MTT)was used to detect cell survival rate.Relevant kits were used to detect the levels of lactate de-hydrogenase(LDH),creatine kinase isoenzyme(CK-MB),Cardiac troponin(cTnI),Malondiadehyde(MDA),superoxide dismutase(SOD)and Glutathione peroxidase(GSH-Px).Flow cytometry was used to detect cell apoptosis rate and mitochondrial membrane poten-tial.Western blot was used to detect the protein expression of B-lymphocytoma-2-associated X protein(Bax),B-lymphocytoma-2(Bcl-2)and cleaved aspartate-specific cysteine proteinase-3(cleaved Caspase-3).Results Clozapine significantly increased circHIPK3,LDH,CK-MB,cTnI,MDA,apoptosis rate,Bax,cleaved Caspase-3 levels of H9C2 cells(P＜0.05),and significantly decreased cell survi-val rate,SOD,GSH-Px,Bcl-2 levels and mitochondrial membrane potential of H9C2 cells(P＜0.05).CircHIPK3 overexpression promot-ed the effect of clozapine on the above indexes(P＜0.05),while silencing circHIPK3 reversed the effect of clozapine on the above indexes(P＜0.05).Conclusion Clozapine can induce H9C2 myocardial cells injury by promoting circHIPK3 expression,and the mecha-nism may be related to the activation of oxidative stress and mitochondrial apoptosis pathways by circHIPK3.
万方数据
维普
