miR-19a-3p对缺血再灌注心肌细胞焦亡的影响

Effect of miR-19a-3p on Myocardial Pyroptosis after Ischemia-Reperfusion

目的 探究miR-19a-3p在心肌缺血再灌注过程中对心肌细胞焦亡情况的影响。方法 动物实验方面,将小鼠分为对照组、模型组、过表达组、抑制组。对照组小鼠仅接受开胸、关胸手术处理;模型组、过表达组、抑制组通过开胸手术方式建立心肌缺血再灌注动物模型,在再灌注同时分别使用腺病毒转染空载腺病毒、miR-19a-3p类似物、miR-19a-3p抑制剂转染并建立模型,模型建立后通过超声心动图、Masson染色、TUNEL染色、TTC染色评估小鼠心脏功能、心肌纤维化、心肌细胞焦亡、心肌梗死情况,并使用Western blot检测各组小鼠焦亡相关蛋白和IRF-8/MAPK信号通路相关蛋白表达情况。细胞实验方面,另取胎鼠原代心肌细胞作为研究对象,将胎鼠原代心肌细胞分为对照组、模型组、过表达组、抑制组。对照组小鼠在正常培养环境中培养且不接受腺病毒转染处理;模型组、过表达组、抑制组分别接受腺病毒转染空载腺病毒、miR-19a-3p类似物、miR-19a-3p抑制剂转染,然后在缺氧/复氧条件下培养。对各组细胞进行TUNEL染色,并使用Western blot检测焦亡相关蛋白和IRF-8/MAPK信号通路相关蛋白。结果 在动物实验中,过表达组和对照组小鼠心脏射血分数显著高于模型组,抑制组小鼠射血分数显著低于模型组;过表达组和对照组小鼠心肌梗死体积、纤维化面积显著低于模型组,抑制组小鼠心肌梗死体积和纤维化面积显著高于模型组。在动物和细胞实验中,过表达组和对照组焦亡细胞比例均显著低于模型组,抑制组焦亡细胞比例均显著高于模型组;过表达组和对照组细胞焦亡相关蛋白表达量显著低于模型组,抑制组细胞焦亡相关蛋白显著高于模型组;过表达组和对照组细胞IRF-8/MAPK信号通路相关蛋白表达量显著低于模型组,抑制组IRF-8/MAPK信号通路相关蛋白表达量显著高于模型组。结论miR-19a-3能够通过抑制IRF-8-MAPK信号通路抑制缺氧/复氧心肌细胞焦亡。

Objective To investigate the effect of mir-19a-3p on myocardial pyroptosis after myocardial ischemiareperfusion.Methods In terms of animal experiments,mice were divided into control group,model group,overexpression group and inhibition group.The mice in the control group only received open chest surgery and closed chest surgery.An animal model of myocardial ischemia/reperfusion was established by transthoracic surgery in the model group,overexpression group and inhibition group.At the same time of reperfusion,adenovirus was used to transfect empty adenovirus,miR-19a-3p analog and miR-19a-3p inhibitor respectively.Then the mouse heart function,myocardial fibrosis,cardiomyocytes pyroptosis and myocardial infarction were evaluated by echocardiography,Masson staining,TUNEL staining and TTC staining,WB was used to detect the expression of pyroptosis related protein and IRF-8/MAPK signal pathway related protein in mice of each group.In terms of cell experiment,the primary fetal rat cardiomyocytes were taken as the research object and divided into control group,model group,overexpression group and inhibition group.The cells in the control group were cultured in normal culture environment without adenovirus transfection.The cells in model group,overexpression group and inhibition group were transfected with adenovirus,miR-19a-3p analog and miR-19a-3p inhibitor respectively,and then cultured under hypoxia/reoxygenation conditions.Then TUNEL staining was performed on cells of each group,and Western blot was used to detect pyroptosis related proteins and IRF-8/MAPK signal pathway related proteins.Results In animal experiments,the heart ejection fraction of mice in overexpression group and control group was significantly higher than that in model group,and that of mice in inhibition group was significantly lower than that in model group.The myocardial infarct volume and fibrosis area of mice in overexpression group and control group were significantly lower than those in model group,and the myocardial infarct volume and fibrosis area of mice in inhibition group were significantly higher than those in model group.Both in animal and cell experiments,the proportion of pyroptosis dead cells in over-expression group and the control group was significantly lower than that in model group,and the proportion of pyroptosis cells in inhibition group was significantly higher than that in model group.The expression of pyroptosis related proteins in over-expression group and the control group was significantly lower than that in model group,and that in inhibition group was significantly higher than that in model group.The expression of IRF-8/MAPK signal pathway related proteins in over-expression group and the control group was significantly lower than that in model group,and the expression of IRF-8/MAPK signal pathway related proteins in inhibition group was significantly higher than that in model group.Conclusions mir-19a-3 can inhibit the pyroptosis of the cardiomyocytes in the condition of hypoxic/reoxygenation by inhibiting IRF-8-mapk signaling pathway.