摘要
目的:探究环状RNA Lrp6(circLrp6)对过氧化氢(H_(2)O_(2))诱导的H9c2心肌细胞凋亡的影响。方法:通过Sanger测序和RNase R酶切验证circLrp6的环状结构;细胞荧光原位杂交分析circLrp6的亚细胞定位;通过RT-qPCR检测H_(2)O_(2)处理的H9c2细胞和缺血再灌注损伤的小鼠心脏组织中circLrp6的差异表达水平;采用TUNEL染色检测circLrp6是否影响H_(2)O_(2)诱导的心肌细胞凋亡水平;通过生物信息学的方法预测与circLrp6相互结合的下游靶点及其结合位点。结果:circLrp6具有环状结构,定位于细胞核,它在H_(2)O_(2)处理后的H9c2心肌细胞和缺血再灌注的心脏组织中表达水平下调(P<0.05),过表达circLrp6对H_(2)O_(2)处理的心肌细胞具有保护作用,表现为TUNEL染色阳性率下降(P<0.05)。另外,circLrp6具有结合miRNA和蛋白质的潜能。结论:circLrp6对H_(2)O_(2)诱导的心肌细胞凋亡具有抑制作用。
AIM:To investigate the effect of circular RNA Lrp6(circLrp6)on hydrogen peroxide(H_(2)O_(2))-induced cardiomyocyte apoptosis.METHODS:The circular structure of circLrp6 was verified by Sanger sequencing and RNase R digestion.The subcellular localization of circLrp6 was detected by fluorescence in situ hybridization(FISH).RT-qPCR was used to detect whether H_(2)O_(2) and myocardial ischemia-reperfusion influenced the expression of circLrp6 in cardiomyocytes.TUNEL staining was used to detect the apoptosis of cardiomyocytes.The downstream targets and binding sites of circLrp6 were predicted by bioinformatic methods.RESULTS:Circularly structural circLrp6 was localized in the nucleus.circLrp6 was down-regulated after treated with H_(2)O_(2) in H9c2 cells(P<0.05).The expression of circLrp6 was also decreased in cardiac tissues of the mice with myocardial ischemia-reperfusion.TUNEL staining showed that overexpression of circLrp6 attenuated the apoptosis of H9c2 cells induced by H_(2)O_(2).In addition,circLrp6 has the potential to combine with miRNA and protein.CONCLUSION:circLrp6 inhibits H_(2)O_(2)-induced cardiomyocyte apoptosis.
作者
丁林
李萌阳
王梦宇
法鸿鸽
房芯羽
王建勋
DING Lin;LI Meng-yang;WANG Meng-yu;FA Hong-ge;FANG Xin-yu;WANG Jian-xun(School of Basic Medical Sciences,Qingdao University,Qingdao 266021,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第3期412-419,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81900259)。
