摘要
目的研究miR-208a调控QKI5表达对大鼠心肌缺血再灌注损伤的影响及作用机制。方法将雄性SD大鼠32只随机选择分为假手术组(Sham组)、缺血再灌注组(I/R组)、I/R+antagomir阴性对照组(I/R+antamiR-NC组)和I/R+miR-208a antagomir组(I/R+anta-miR-208a组),构建大鼠心肌I/R模型后检测各组大鼠心脏左室射血分数(LVEF)、左室短轴缩短率(LVFS)和左室每搏量(SV);ELISA法检测血清肌酐激酶同工酶(CKMB)和心肌肌钙蛋白(cTnI)以及IL-1β、IL-6和肿瘤坏死因子(TNF-α)水平;采用HE染色观察各组心肌组织病理变化,采用TUNEL染色检测心肌组织细胞凋亡;RT-PCR检测心肌组织miR-208a和QKI5 mRNA表达,Western blot检测心肌组织QKI5蛋白和凋亡蛋白Caspase-3的相对表达水平。结果与Sham组比较,I/R组中LVEF、LVFS、SV值降低,心肌细胞凋亡率升高,血清中CK-MB、cTnI和IL-1β、IL-6、TNF-α水平及心肌组织中miR-208a和Caspase-3表达升高,而QKI5表达水平降低(P<0.05);与I/R+anta-miR-NC组比较,I/R+anta-miR-208a组中LVEF、LVFS、SV值升高,心肌细胞凋亡率降低,血清中CK-MB、cTnI和IL-1β、IL-6、TNF-α及心肌组织中miR-208a和Caspase-3表达水平降低,而QKI5表达升高(P<0.05)。相关性分析发现miR-208a与QKI5表达呈负相关。结论抑制miR-208a可能通过调控QKI5表达减轻心肌缺血再灌注损伤中炎症反应,发挥心肌保护作用。
Objective To investigate the effect and mechanism of miR-208a on myocardial injury induced by myocardial ischemia reperfusion by regulating the expression of QKI5 in rats.Methods Thirty-two male SD rats were randomly divided into sham group,I/R group,I/R+anta-miR-NC group and I/R+anta-miR-208a group.The rat model of myocardial I/R was established,and the LVEF,LVFS and SV were detected in each group.The levels of CK-MB,cTnI,IL-1β,IL-6,TNF-αwere measured by ELISA.HE staining was used to observe the pathological changes of myocardial tissue in each group,and TUNEL staining was used to detect myocardial cell apoptosis.RT-PCR was used to detect the expression of miR-208a and QKI5 mRNA in myocardial tissue,and Western blot was used to detect the realative expression levels of QKI5 protein and Caspase-3 in myocardial tissue.Results Compared with the Sham group,the levels of LVEF,LVFS,SV in the I/R group were decreased,while the apoptosis rate of cardiomyocytes was increased,and the levels of CK-MB,cTnI,IL-1β,IL-6,TNF-αin serum and the expressions of miR-208a and Caspase-3 in the myocardial tissue were increased,and the expressions of QKI5 were decreased(P<0.05).Compared with the I/R+anta-miR-NC group,the levels of LVEF,LVFS,SV in the I/R+anta-miR-208a group were increased,while the apoptosis rate of cardiomyocytes was decreased,and the levels of CK-MB,cTnI,IL-1β,IL-6,TNF-αin serum and the expressions of miR-208a and Caspase-3 in the myocardial tissue were decreased,and the expressions of QKI5 were increased(P<0.05).Correlation analysis found that the expression of miR-208a and QKI5 had negative correlation in the I/R rats.Conclusion Inhibition of miR-208a may alleviate the inflammatory response to exert myocardial protective function in myocardial ischemia-reperfusion injury by regulating the expression of QKI5.
作者
杨伟
陈洪艳
陈文栋
王燕琼
白向锋
YANG Wei;CHEN Hongyan;CHEN Wendong;WANG Yanqiong;BAI Xiangfeng(Dept.of Anesthesiology,The 1st Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China;Dept.of Ultrasound,The 1st Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China;Dept.of Cardiovascular Surgery,The 1st Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China)
出处
《昆明医科大学学报》
CAS
2022年第7期18-24,共7页
Journal of Kunming Medical University
基金
云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目[2019FE001(-045)]
云南省教育厅科学研究基金资助项目[2021J0230]。
