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目的:探讨上调miRNA378*表达对柯萨奇B3病毒(CVB3)感染心肌细胞凋亡、网腔钙结合蛋白、内质网应激及信号通路因子的作用。方法:实验分4组:对照组(正常细胞)、CVB3感染组(正常细胞+CVB3)、miRNA378*过表达对照组(正常细胞+CVB3+转染miRNA378*空表达质粒)、miRNA378*过表达组(正常细胞+CVB3+转染miRNA378*过表达质粒)。原代培养乳鼠心肌细胞,采用免疫组织化学方法检测培养乳鼠心室肌细胞α-SMA蛋白,慢病毒质粒转染心室肌细胞,除对照组外,其他各组心肌细胞感染CVB3,采用TUNEL技术检测各组心肌细胞凋亡率;用Western blotting技术检测各组心肌细胞网腔钙结合蛋白、内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PERK、P-PERK、eIF2α、ATF4、CHOP表达。结果:与CVB3感染组比较,miRNA378*过表达组心肌细胞凋亡率明显减少,网腔钙结合蛋白表达增加,而GRP78、P-PERK、eIF2α、ATF4、CHOP表达均减少(均P<0.01),PERK表达差异无统计学意义。结论:上调CVB3感染心肌细胞miRNA378*表达可引起心肌细胞凋亡减少,网腔钙结合蛋白表达增多,进而缓解内质网应激,并抑制内质网应激凋亡信号通路因子表达。
AIM: To investigate the effects of up-regulation miRNA378 * on CVB3-induced cardiomyocyte apoptosis, Ca2 +, endoplasmic reticulum stress and signal pathway factors. Methods: The experiment was divided into 4 groups: control group (normal cells), CVB3 infected group (normal cells + CVB3), miRNA378 * overexpression control group (normal cells + CVB3 + transfected miRNA378 * empty plasmid), miRNA378 * Normal cells + CVB3 + transfected miRNA378 * overexpression plasmid). Primary cultured neonatal rat cardiomyocytes were detected by immunohistochemical method cultured neonatal rat ventricular myocytes α-SMA protein, lentiviral plasmid transfected ventricular myocytes, in addition to the control group, the other groups CVB3 myocardial cells infected with TUNEL technology The apoptosis rate of cardiomyocytes in each group was detected by Western blotting. The expressions of ERK, P-PERK, eIF2α, ATF4 and ERK were detected by Western blotting. , CHOP expression. Results: Compared with CVB3 infection group, the apoptosis rate of myocardial cells in miRNA378 * overexpression group was significantly decreased, and the expression of Ca2 +, but GRP78, P-PERK, eIF2α, ATF4 and CHOP were decreased (all P <0.01) There was no significant difference in PERK expression. CONCLUSION: Up-regulation of miRNA378 * expression in CVB3-infected cardiomyocytes can reduce the apoptosis of cardiomyocytes and increase the expression of calcium-binding protein in the cytoplasm, thereby relieving the ER stress and inhibiting the expression of ER signaling pathway.