微小RNA-98-5p靶向Kruppel样转录因子9对大鼠心肌缺血/再灌注损伤的保护作用  

Protective effects of micro RNA-98-5p targeting Kruppel-like factor 9 against myocardial ischemia-reperfusion injury in rats

作　　者：卢芬[1] 杨晴[2] 邵文君[3] 孙振坤 王恩漫[5] 张伟娜 LU Fen;YANG Qing;SHAO Wen-jun;SUN Zhen-kun;WANG En-man;ZHANG Wei-na(Department of Internal Medicine,Shangqiu Medical College,He'nan Shangqiu 476000,China;Department of Obstetrics and Gynecology,Shangqiu Medical College,He'nan Shangqiu 476000,China;He1 nan Provincial People's Hospital,Zhengzhou 450000,China;Department of Internal Medicine,Shangqiu Municipal Hospital,He'nan Shangqiu 476000,China;Department of Surgery,Shangqiu Medical College,He'nan Shangqiu 476000,China;School of Pharmacy,Shangqiu Medical College,He'nan Shangqiu 476000,China)

机构地区：[1]商丘医学高等专科学校内科,河南商丘476000 [2]商丘医学高等专科学校妇产科,河南商丘476000 [3]河南省人民医院神经内科,郑州450000 [4]商丘市立医院内科,河南商丘476000 [5]商丘医学高等专科学校外科,河南商丘476000 [6]商丘医学高等专科学校药学院,河南商丘476000

出　　处：《解剖学报》2022年第3期347-353,共7页Acta Anatomica Sinica

基　　金：河南省高等学校重点科研项目(20B150018);商丘医学高等专科学校青年科研基金(科技攻关2020-Qnjj-002);商丘医学高等专科学校博士科研启动项目(BSJH001)。

摘　　要：目的探讨微小RNA(miR)-98-5p靶向Kruppel样转录因子9(KLF9)对大鼠心肌缺血/再灌注(MI/R)损伤的保护作用。方法50只大鼠随机分为假手术组、模型组、miR-98-5p agomir组、agomir-NC组及miR-98-5p agomir+pcDNA-3.1-KLF9组,每组10只。通过冠状动脉结扎法建立MI/R注模型。HE染色观察心肌组织病理情况;TUNEL检测心肌组织细胞凋亡情况;ELISA检测血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)含量;Real-time PCR检测心肌组织miR-98-5p、KLF9 mRNA表达水平;Western blotting检测心肌组织KLF9、Bax和JAK2/STAT3信号通路相关蛋白表达;双荧光素酶报告实验验证miR-98-5p与KLF9的关系。结果与假手术组相比,模型组大鼠心肌细胞排列较乱,出现坏死;心肌组织细胞凋亡率、血清CK、CK-MB、LDH含量均升高,心肌组织miR-98-5p表达水平下降,KLF9 mRNA和蛋白及p-JAK2和p-STAT3蛋白表达均升高(P<0.05)。过表达miR-98-5p后,大鼠心肌细胞排列较为整齐,心肌细胞坏死减少;心肌组织细胞凋亡率、血清CK、CK-MB、LDH含量及心肌组织p-JAK2、p-STAT3蛋白表达均下降(P<0.05)。双荧光素酶报告实验结果验证KLF9是miR-98-5p的靶基因。过表达KLF9逆转了miR-98-5p agomir对心肌损伤大鼠产生的作用。结论MiR-98-5p靶向KLF9改善MI/R大鼠心肌损伤,其机制可能与miR-98-5p调控JAK2/STAT3信号通路抑制心肌细胞凋亡相关。Objective To investigate the protective effect of micro RNA(miR)-98-5 p targeting Kruppel-like factor 9(KLF9)against myocardial ischemia-reperfusion(MI/R)injury in rats.Methods Totally 50 rats were randomly divided into sham operation group,model group,miR-98-5 p agomir group,agomir-NC group,and miR-98-5 p agomir+pcDNA-3.1-KLF9 group,10 in each group.MI/R model was established by coronary artery ligation.The pathological changes of myocardial tissue were detected by HE staining.The myocardial apoptosis were detected by TUNEL.The levels of creatine kinase(CK),creatine kinase isoenzymes(CK-MB)and lactate dehydrogenase(LDH)in serum were detected by ELISA.The expression levels of miR-98-5 p and KLF9 mRNA were detected by Real-time PCR.The expression of KLF9,Bax and JAK2/STAT3 pathway relative protein were detected by Western blotting.Dual luciferase assay verified the relationship between miR-98-5 p and KLF9.Results Compared with the sham operation group,the arrangement of myocardial cells in the model group was disordered,and the myocardial cells appeared necrosis.The apoptosis rate of myocardial cells,serum CK,CK-MB and LDH contents increased,the expression level of miR-98-5 p decreased,and the expression levels of KLF9 mRNA and protein,p-JAK2 and p-STAT3 protein increased(P<0.05).After the overexpression of miR-98-5 p,the myocardial cells arranged more orderly and the myocardial cell necrosis decreased.The apoptosis rate of myocardial tissue,the contents of CK,CK-MB and LDH in serum and the expression of p-JAK2 and p-STAT3 protein were decreased(P<0.05).The result of dual luciferase assay showed that KLF9 was the target gene of miR-98-5 p.The overexpression of KLF9 reversed the effects of miR-98-5 p agomir on myocardial injury.Conclusion MiR-98-5 p targeting KLF9 can improve the myocardial injury of MI/R rats.The mechanism may be related to the regulation of JAK2/STAT3 signal pathway by miR-98-5 p which inhibit myocardial cell apoptosis.

关 键 词：微小RNA-98-5p Kruppel样转录因子9 心肌缺血/再灌注 Janus激酶2/信号转导子与转录激活子3信号通路 实时定量聚合酶链反应 免疫印迹法 大鼠 

分 类 号：R542.2[医药卫生—心血管疾病]