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目的:研究阿霉素损伤心肌细胞miRNA378*与网腔钙结合蛋白(calumenin)、内质网应激伴侣蛋白GRP78、凋亡因子bax及bcl-2相关性。方法:实验分6组:对照组、阿霉素组、miRNA378*过表达对照组、miRNA378*过表达组、miRNA378*沉默对照组、miRNA378*沉默组。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,实时荧光定量PCR技术检测各组心肌细胞miRNA378*、calumenin、GRP78、bax及bcl-2mRNA表达。结果:1与对照组相比较,阿霉素组心肌细胞calumenin mRNA表达明显减少(P<0.01),GRP78 mRNA表达增加(P<0.01),bax mRNA表达增加(P<0.01),bcl-2mRNA表达明显减少(P<0.01)。2与阿霉素组相比较,miRNA378*过表达组心肌细胞calumenin mRNA表达增加(P<0.01),bcl-2mRNA表达增加(P<0.01),而GRP78mRNA表达减少(P<0.01),bax mRNA表达减少(P<0.01)。与阿霉素组相比较,miRNA378*沉默组GRP78、bax mRNA表达增加(P<0.01),bcl-2mRNA表达减少(P<0.01)。结论:阿霉素损伤可能引起心肌细胞calumenin表达减少进而发生内质网应激;上调miRNA378*表达会增加阿霉素损伤心肌细胞calumenin表达缓解内质网应激,进而抑制心肌细胞凋亡;而沉默miRNA378加重阿霉素损伤心肌细胞内质网应激及促进心肌细胞凋亡。
Objective: To study the relationship between miRNA378 * and calumenin, endoplasmic reticulum stress chaperone GRP78, bax and bcl-2 in adriamycin-injured myocardial cells. Methods: The experiment was divided into 6 groups: control group, doxorubicin group, miRNA378 * overexpression control group, miRNA378 * overexpression group, miRNA378 * silencing control group and miRNA378 * silencing group. The primary cultured neonatal rat cardiomyocytes were cultured and the α-SMA protein was detected by immunohistochemistry. The lentiviral plasmids were transfected into ventricular myocytes. Real-time fluorescent quantitative PCR was used to detect the expression of miRNA378 *, calumenin, GRP78 , Bax and bcl-2 mRNA expression. Compared with the control group, the expression of calumenin mRNA in adriamycin group was significantly decreased (P <0.01), GRP78 mRNA expression was increased (P <0.01), bax mRNA expression was increased (P <0.01) Significantly reduced (P <0.01). Compared with doxorubicin group, the expression of calumenin mRNA in the miRNA378 * overexpression group increased (P <0.01), the expression of bcl-2 mRNA increased (P <0.01), the expression of GRP78 mRNA decreased (P <0.01) Decrease (P <0.01). Compared with doxorubicin group, miRNA378 * silencing group increased GRP78 and bax mRNA expression (P <0.01) and decreased bcl-2 mRNA expression (P <0.01). CONCLUSION: Doxorubicin injury may lead to the decrease of calumenin expression in cardiomyocytes and the occurrence of endoplasmic reticulum stress. Upregulation of miRNA378 * may increase the expression of calumenin in cardiomyocytes and reduce the endoplasmic reticulum stress, thereby inhibiting cardiomyocyte apoptosis. Silencing miRNA378 aggravates adriamycin-induced cardiomyocyte endoplasmic reticulum stress and promotes cardiomyocyte apoptosis.