脂联素依赖p38-MAPK途径抑制衣霉素诱导的内质网应激致心肌细胞凋亡

发布时间：2018-01-15 11:45

本文关键词：脂联素依赖p38-MAPK途径抑制衣霉素诱导的内质网应激致心肌细胞凋亡　出处：《中国动脉硬化杂志》2012年07期 　论文类型：期刊论文

【摘要】：目的通过原代培养SD大鼠的乳鼠心肌细胞建立衣霉素心肌细胞内质网应激损伤模型,观察脂联素对心肌细胞内质网应激致细胞凋亡的作用及其机制。方法采用酶消化法原代培养乳鼠心肌细胞,倒置相差显微镜下观察细胞生长,通过α-肌动蛋白免疫荧光法对培养的心肌细胞进行鉴定。选用原代培养3～4天的心肌细胞,随机分为五组:对照组、1 mg/L衣霉素组、1 mg/L衣霉素+100 mg/L脂联素组、1 mg/L衣霉素+3μmol/LSB203580组及1 mg/L衣霉素+3μmol/L SB203580+100 mg/L脂联素组。实验终止后,在倒置相差显微镜下观察心肌细胞形态变化,通过流式细胞术检测心肌细胞凋亡,用qRT-PCR及免疫荧光法检测内质网应激指标GRP78和CHOP的mRNA及蛋白表达。结果与对照组相比,给予衣霉素后,细胞凋亡率显著增加,GRP78和CHOP的mR-NA及蛋白表达增加。脂联素预处理后给予衣霉素,可较大程度地逆转上述指标变化,细胞凋亡率显著下降,GRP78和CHOP的mRNA及蛋白表达减少;而加用p38-MAPK抑制剂SB203580后脂联素的保护作用明显减弱,凋亡率显著增加,GRP78和CHOP的mRNA及蛋白表达增高,但较单纯衣霉素处理组凋亡率低,GRP78和CHOP的mRNA及蛋白表达也减少。结论衣霉素可使GRP78和CHOP表达增强,启动内质网应激,导致心肌细胞凋亡,脂联素可以通过减轻内质网应激逆转衣霉素所致的心肌细胞凋亡作用,对心肌细胞有保护作用,且这种保护作用部分是通过p38-MAPK途径实现的。
[Abstract]:Objective to establish a model of endoplasmic reticulum (ER) stress injury in neonatal rat cardiomyocytes by primary culture of Sprague-Dawley (SD) rats. To observe the effect of adiponectin on apoptosis induced by endoplasmic reticulum stress in cardiomyocytes and its mechanism. Methods the primary cultured neonatal rat cardiomyocytes were cultured by enzyme digestion and the cell growth was observed under inverted phase contrast microscope. The cultured cardiomyocytes were identified by 伪 -actin immunofluorescence. The cultured cardiomyocytes were divided into five groups randomly: control group (1 mg/L). 1 mg/L itamycin 100 mg/L adiponectin group. 1 mg/L chlortetracycline 3 渭 mol/LSB203580 group and 1 mg/L chlortetracycline 3 渭 mol/LSB203580 100. Mg/L adiponectin group. The morphologic changes of cardiomyocytes were observed under inverted phase contrast microscope and apoptosis of cardiomyocytes was detected by flow cytometry. The expression of mRNA and protein in endoplasmic reticulum stress index (GRP78 and CHOP) was detected by qRT-PCR and immunofluorescence. The expression of mR-NA and protein in GRP78 and CHOP increased. After pretreatment with adiponectin, the changes of the above indexes were reversed to a large extent, and the apoptosis rate decreased significantly. The expression of mRNA and protein in GRP78 and CHOP was decreased. However, the protective effect of adiponectin on p38-MAPK inhibitor SB2035 80 was significantly weakened, and the expression of mRNA and protein in GRP78 and CHOP were significantly increased after adding p38-MAPK inhibitor SB2035 80. However, the expression of mRNA and protein in GRP78 and CHOP was lower than that in the control group. Conclusion the expression of GRP78 and CHOP was increased and endoplasmic reticulum stress was induced by chlortetracycline. Adiponectin can reverse the apoptosis of cardiomyocytes induced by itamycin by reducing endoplasmic reticulum stress and has protective effect on cardiomyocytes. This protective effect is partly achieved by p38-MAPK pathway.
【作者单位】：山西医科大学第二医院心内科;
【基金】：国家自然科学基金(81170198)
【分类号】：R363
【正文快照】： 内质网是哺乳动物细胞中一种重要的亚细胞器。膜P分泌性蛋白、氨基多糖、胆固醇及钙信号等的代谢均与内质网功能直接相关,如分泌性蛋白的合成与空间折叠、蛋白质糖基化修饰、蛋白质分泌等均在内质网内发生。目前研究发现,心肌细胞内质网功能障碍可能是心肌缺血再灌注损伤的重

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