摘要
目的建立小鼠急性心肌缺血动物模型,探讨急性心肌缺血中纤溶酶原激活物抑制剂-1(PAI-1)的变化以及微小RNA(miRNA,miR)-30b的作用及机制.方法小鼠分成模型组和对照组,模型组气管插管,呼吸机辅助呼吸,胸骨左缘3~4肋间横切口,显露心脏,以6-0无损伤缝合线缝扎前降支.观察小鼠缺血前后心电图J点电压变化.取血清检测过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性.模型组和对照组处死小鼠前抽血,处死小鼠后取心肌组织.以实时定量反转录聚合酶链反应(RT-qPCR)、蛋白质印迹法(Western blot)、酶联免疫吸附试验(ELISA)技术检测小鼠心肌和血液中的PAI-1和miR-30b表达变化.应用SPSS 18.0统计软件分析;多组计量资料采用One-way ANOVA,方差齐时采用LSD和SNK法,方差不齐时采用Tamhane's T2或Dunnett's T3法.结果模型组大鼠血液中PAI-1的mRNA和蛋白表达均明显上调(=1.031,P<0.05),心肌中PAI-1的mRNA和蛋白表达上调(t=-4.573,P<0.05),miR-30b表达均明显下调,差异有统计学意义(t=-1.968,P<0.05);双荧光素酶基因报告表明PAI-1为miR-30b直接靶基因.结论急性心肌缺血模型中PAI-1表达明显上调,出现纤溶亢进、增强,可能与miR-30b下调有关,miR-30b可能通过对PAI-1调控参与急性心肌缺血过程.
Objective Through the establishment of an animal model of acute myocardial ischemia in mice,to investigate the changes of plasminogen activator inhibitor-1(PAI-1)and the action mechanism of microRNA(miRNA,miR)-30b during acute myocardial ischemia.Methods Mice were divided into the model group and the control group.Acute myocardial ischemia animal model was established after occlusion of left anterior descending coronary artery in the model group.The changes of PAI-1 and miR-30b in the myocardium and blood of mice were detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR),Western blotting,and enzyme linked immunosorbent assay(ELISA).Results The mRNA and protein expression levels of PAI-1 in the myocardium and blood of the model group were significantly up-regulated,and the miR-30b expression was significantly down-regulated,with statistically significant differences(t=1.031,-4.573,-1.968,P<0.05).The dual luciferase gene report indicated that PAI-1 was a direct target gene of miR-30b.Conclusion The expression of PAI-1 in the model of the acute myocardial ischemia was significantly up-regulated and hyperfibrinolysis occurred,which may be related to the down-regulation of miR-30b.The miR-30b may be involved in the process of acute myocardial ischemia through the regulation of PAI-1.
作者
李斌
胡杰
陈兴澎
赵勇鹏
权晓强
Li Bin;Hu Jie;Chen Xinpeng;Zhao Yongpeng;Quan Xiaoqiang(Department of Cardiac Surgery,Zhengzhou University Affiliated Central Hospital of Luoyang,Luoyang 471009,China;Department of Cardiac Surgery,Henan Provincial People’s Hospital/Fuwai Central China Cardiovascular Hospital/People’s Hospital of Zhengzhou University,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第9期1567-1570,共4页
Chinese Journal of Experimental Surgery
基金
河南省洛阳市科技局基金项目(1820007A).
