摘要
目的探究微小RNA-22(miR-22)通过靶向调控Notch通路对冠状动脉粥样硬化性心脏病(冠心病,CHD)动物模型免疫炎症反应的影响。方法通过高脂饮食构建CHD大鼠模型,qPCR或Western blot检测冠状动脉(冠脉)组织中miR-22和Notch通路的变化。构建原代大鼠冠脉内皮细胞(CAECs)模型,通过双荧光素酶报告在CAECs中验证miR-22靶向Jagged1(JAG1)。将CAECs分为NC组、mimic组、mimic+JAG1组和JAG1组,共4组。通过CCK-8、Hoechst 33258染色和ELISA检测细胞生长、凋亡和培养基中炎性因子水平。结果与对照组比较,CHD组大鼠冠脉中miR-22显著下调,而Notch通路显著上调(P<0.05)。miR-22与JAG1 mRNA靶向结合。mimic组的miR-22和JAG1组的JAG1 mRNA显著高于NC组(P<0.05)。mimic组的JAG1 mRNA水平显著低于NC组(P<0.05)。mimic+JAG1组的JAG1 mRNA水平显著高于mimic组(P<0.05)。结论miR-22可能通过靶向抑制Notch通路抑制CHD中CAECs的凋亡,并抑制炎性细胞因子分泌。
Objective To study the influence of microRNA-22(miR-22)on immune inflammatory response in animal model of coronary heart disease(CHD)through targeted regulation of Notch pathway.Methods The rat model of CHD was established by high-fat diet,and changes of miR-22 and Notch in coronary artery tissue were detected by using qPCR and Western blotting assay.The model of coronary artery endothelial cells(CAECs)was established in primary rats,and miR-22 targeting Jagged1(JAG1)in CAECs was verified by using dual-luciferase reporter.CAECs were divided into NC group,mimic group,mimic+JAG1 group and JAG1 group.The growth and apoptosis of CAECs and levels of inflammatory factors in culture medium were detected by using CCK-8 and ELISA after Hoechst 33258 staining.Results Compared with control group,miR-22 was significantly down-regulated and Notch pathway was significantly up-regulated in CHD group(P<0.05).MiR-22 and JAG1 mRNA showed a target integration.MiR-22 in mimic group and JAG1 mRNA in JAG1 group were significantly higher that those in NC group(P<0.05).The level of JAG1 mRNA was significantly lower in mimic group than that in NC group(P<0.05).The level of JAG1 mRNA was significantly higher in mimic+JAG1 group than that in mimic group(P<0.05).Conclusion MiR-22 may inhibit the apoptosis of CAECs and secretion of inflammatory cytokines in CHD through target inhibition of Notch pathway.
作者
张洪博
赵宇
陶亮
李春峰
Zhang Hongbo;Zhao Yu;Tao Liang;Li Chunfeng(First Department of Cardiovascular Medicine,Second Affiliated Hospital of Qiqihar Medical College,Qiqihar 161000,China;不详)
出处
《中国循证心血管医学杂志》
2020年第8期959-962,共4页
Chinese Journal of Evidence-Based Cardiovascular Medicine
基金
齐齐哈尔市科技公关项目(2019L368)。
