CircRNA-32011调控三氧化二砷诱导的心肌细胞凋亡

CircRNA-32011 regulates apoptosis induced by arsenic trioxide in cardiac myocytes

目的探究环状RNA-32011(CircRNA-32011)在三氧化二砷(arsenic trioxide,ATO)诱导心肌细胞凋亡中的作用。方法采用ATO(终浓度10μmol·L^(-1))孵育乳鼠原代心肌细胞24 h,应用MTT法检测心肌细胞存活率;RT-PCR检测circRNA-32011及凋亡调控蛋白Bcl-2和Bax的mRNA表达水平,计算Bcl-2/Bax;Western blot检测Bcl-2和Bax蛋白表达并计算Bcl-2/Bax。应用过表达质粒和siRNA分别过表达及敲减circRNA-32011验证其在ATO诱导的心肌细胞凋亡中的功能。结果ATO处理后,心肌细胞存活率明显下降,Bcl-2/Bax表达降低,CircRNA-32011表达下调;在ATO处理的心肌细胞中过表达circRNA-32011可明显提高心肌细胞存活率和Bcl-2/Bax表达;敲减circRNA-32011可进一步降低由ATO诱导的心肌细胞存活率和Bcl-2/Bax表达下调。结论CircRNA-32011抑制ATO诱导的心肌细胞凋亡,这将为ATO诱发的心肌损伤提供一种新的潜在治疗策略。

Aim To investigate the effect of circRNA-32011on myocardial apoptosis induced by arsenic triox-ide(ATO).Methods Primary cardiomyocytes of suckling neonate mouse were treated with ATO(final concentration 10μmol·L^(-1))for 24 h.Then cell viability was measured by MTT assay.The mRNA expression levels of Bcl-2/Bax and circRNA-32011 were detected by RT-PCR.Bcl-2/Bax protein expression levels were detected by Western blot.Overexpression and knock down circRNA-32011 respectively by plasmid and siRNA were used to verify its function in ATO-induced cardiomyocyte apoptosis.Results Myocardial cell viability decreased,Bcl-2 expression significantly decreased while Bax expression increased in ATO group compared with the control group.CircRNA-32011 was down-regulated in ATO incubated cardiomyocytes.Overexpression of circRNA-32011 in ATO-incubated cardiomyocytes increased myocardial cell viability and Bcl-2 expression and decreased the expression of Bax.Knockdown of circRNA-32011 could further reduce cardiomyocyte activity and Bcl-2 expression and increase the experssion of Bax induced by ATO.Conclusions CircRNA-32011 protects cardiac myocytes from apoptosis induced by arsenic trioxide,which may provide a new potential therapeutic strategy for ATO-induced myocardial injury.