摘要
目的:探究miR-499-5p在大鼠心肌梗死后心肌细胞凋亡中的作用和相关机制。方法:80只大鼠随机分为假手术组、心肌梗死组(AMI组)、AMI+miR-499-5p mimics组(干预组)、AMI+scramble-NC组(阴性对照组)。TUNEL法检测心肌细胞凋亡;qRT-PCR检测Sox6 mRNA表达;Western blot分析Sox6、Bax、Bcl-xl蛋白表达;超声心动图评估心功能;荧光素酶报告实验检测miR-499-5p是否靶向调控Sox6 3’非编码区。结果:与假手术组比较,AMI组心肌细胞凋亡率增高,Sox6 mRNA和蛋白及Bax蛋白表达增高,Bcl-xl蛋白表达降低,EF值和FS值均降低(P均<0.05);与阴性对照组、AMI组比较,干预组心肌细胞凋亡率、Sox6 mRNA和蛋白及Bax蛋白表达降低,Bcl-xl蛋白表达增高,EF、FS值增加(P均<0.05)。荧光素酶报告实验提示miR-499-5p特异性调控Sox6 3’末端非编码序列。结论:miR-499-5p可能通过靶向抑制Sox6的表达来抑制心肌细胞凋亡,最终改善心功能。
Aim： To investigate the role of miR-499-5 p in cardiomyocyte apoptosis after myocardial infarction in rats and explore its molecular mechanism. Methods： The rats were randomly allocated into sham operation group,AMI group,intervention group（ AMI ＋ intravenous injection of miR-499-5 p mimics after pulse ligation）,and negative control group（AMI ＋ intravenous injection of negative control sequence）. Cardiomyocyte apoptosis was detected by TUNEL staining.Sox6 mRNA was detected by qRT-PCR; the expressions of Sox6,Bax,Bcl-xl protein were detected by Western blot; heart function was detected by Vevo 2100. Luciferase reporter assay was used to detect whether miR-499-5 p targeted Sox6 3＇ noncoding region. Results： Compared with the sham operation group,the apoptosis rate,Sox6 and Bax expression increased,while Bcl-xl expression,the values of EF and FS decreased in AMI group（ P〈0. 05）. Compared with the negative control group and AMI group,the apoptosis rate,Sox6,Bax protein decreased,while Bcl-xl expression and the values of EF and FS increased. Luciferase reporter assay showed that miR-499-5 p specifically regulated the 3 ＇ non-coding region of Sox6. Conclusion：miR-499-5 p may inhibit cardiomyocyte apoptosis by targeted inhibition of Sox6 expression,and ultimately improve cardiac function.
作者
吴丽梦
李恩
周小翠
郭小宏
王蒙
赵英杰
汪涛
WU Limeng;LI En;ZHOU Xiaocui;GUO Xiaohong;WANG Meng;ZHAO Yingjie;WANG Tao(Department of Cardiovascular Medicine,the Second Affiliated Hospital,Zhengzhou University,Zheng-zhou 450014;Geriatric Department,the Second Affiliated Hospital,Zhengzhou University,Zhengzhou 450014)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2018年第5期614-618,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省重点科技攻关项目(142102310109
172102310376)
