摘要
目的探讨环状RNA(circRNA)_0068655对心肌细胞HCM凋亡的影响。方法12只雄性大鼠随机分为假手术组(Sham组)和心肌梗死组(MI组),qRT-PCR检测两组心肌组织中circ_0068655和miR-498表达水平,Western blotting检测两组心肌组织中凋亡相关蛋白的表达。将HCM分为空白对照组、circ_0068655敲低组(sh-circ_0068655组),circ_0068655过表达组、低氧刺激组(Hypoxia组)、敲低对照+低氧刺激组(sh-ctrl+hypoxia组)、circ_0068655敲低+低氧刺激组(sh-circ_0068655+hypoxia组),缺氧的HCM分为敲低对照+miR-498-inhibitor-NC组(sh-ctrl+miR-498-inhibitor-NC组)、circ_0068655敲低+miR-498-inhibitor-NC组(sh-circ_0068655+miR-498-inhibitor-NC组)及circ_0068655敲低+miR-498-inhibitor组(sh-circ_0068655+miR-498-inhibitor组),qRT-PCR检测各组HCM中circ_0068655和miR-498表达量,CCK-8、ELISA及Transwell检测细胞活性、凋亡、迁移及侵袭能力。RNA pull-down及双荧光素酶实验分析circ_0068655和miR-498结合关系。结果MI大鼠心肌组织中circ_0068655的表达高于Sham组,miR-498表达低于Sham组,敲低circ_0068655可以改善缺氧诱导的HCM心肌细胞活力,提高心肌细胞迁移及侵袭能力,降低细胞凋亡率及凋亡相关蛋白表达。circ_0068655可结合并负向调节miR-498的表达。miR-498抑制剂可抑制敲低circ_0068655对缺氧心肌细胞保护作用。结论circ_0068655能通过吸附miR-498促进心肌梗死及心肌细胞凋亡。
Aim To investigate the effects of circular_0068655 on the apoptosis of cardiomyocytes.Methods Twelve male rats were randomly divided into Sham group and acute myocardial infarction group(MI group).qRT-PCR was used to detect the expression of circ_0068655 and miR-498,and Western blotting was performed to detect the expression of apoptosis-related proteins in myocardium of two groups of rats.HCM were divided into blank control group,circ_0068655 knockdown group(sh-circ_0068655 group),circ_0068655 overexpression group,hypoxia group(hypoxia group),knockdown control+hypoxia group(sh-ctrl+hypoxia group),circ_0068655 knockdown+hypoxia(sh-circ_0068655+hypoxia group).Hypoxic HCM were divided into knockdown control+miR-498-inhibitor-NC group(sh-ctrl+miR-498-inhibitor-NC group),circ_0068655 knockdown+miR-498-inhibitor-NC group(sh-circ_0068655+miR-498-inhibitor-NCgroup)and circ_0068655 knockdown+miR-498-inhibitor group(sh-circ_0068655+miR-498-inhibitor group).The expression levels of circ_0068655 and miR-498 in HCM were detected by qRT-PCR,and cell activity,apoptosis,migration and invasion were detected by CCK-8,ELISA and Transwell methods in all groups.The binding relationship between circ_0068655 and miR-498 was analyzed by RNA pull-down and dual luciferase assays.Results The expression of circ_0068655 in myocardial tissue of MI rats was higher than that of Sham group,and the expression of miR-498 was lower than that of Sham group.Knockdown of circ_0068655 could improve the viability of HCM cardiomyocytes induced by hypoxia,improve the migration and invasion ability of cardiomyocytes,and reduce the apoptosis rate and apoptosis-related protein expression.circ_0068655 can bind and negatively regulate the expression of miR-498.miR-498-inhibitors can inhibit the protective effects of circ_0068655 knockdown on hypoxic cardiomyocytes.Conclusion circ_0068655 can promote the apoptosis of cardiomyocytes in myocardial infarction by sponging miR-498.
作者
崔冰
柴巧英
李鑫平
王丹丹
陈兰兰
韩继如
张海军
CUI Bing;CHAI Qiaoying;LI Xinping;WANG Dandan;CHEN Lanlan;HAN Jiru;ZHANG Haijun(Department of Clinical Laboratory,Handan First Hospital,Handan 056001,Hebei,China;Department of Cardiology,Handan First Hospital,Handan 056001,Hebei,China;Department of General Medicine,Handan Central Hospital,Handan 056001,Hebei,China)
出处
《中南医学科学杂志》
CAS
2024年第4期539-543,573,共6页
Medical Science Journal of Central South China
基金
河北省医学科学研究课题(20220012,20200438)。
