D-半乳糖诱导的心肌细胞自噬下降与microRNA的表达变化有关

发布时间：2018-06-30 08:49

  本文选题：衰老心肌细胞 + 自噬　； 参考：《中国动脉硬化杂志》2017年04期

【摘要】：目的探讨microRNA(miRNA)和衰老心肌细胞自噬之间的关系。方法使用0、4、8、16、20 g/L D-半乳糖刺激大鼠心肌细胞H9c2衰老,刺激3、6、9天后,β-半乳糖苷酶染色法检测细胞衰老水平;电镜观察D-半乳糖刺激衰老大鼠心肌细胞自噬体形成;分别采用实时荧光定量PCR(qRT-PCR)及蛋白质印迹法(Western blot)检测衰老大鼠心肌细胞与正常H9c2细胞中miRNA-30a、miRNA-126、miRNA-204及自噬蛋白LC3B II/LC3B I表达水平。结果与对照组比较,随着药物浓度的升高及刺激天数的增加,细胞染色逐渐加深,在D-半乳糖浓度为8 g/L刺激9天时,细胞染色最明显;电镜结果显示,D-半乳糖诱导衰老心肌细胞自噬体减少;Western blot结果显示,衰老心肌细胞LC3B II/LC3B I水平降低;实时荧光定量PCR结果显示,诱导衰老组心肌细胞中,miRNA-30a、miRNA-126表达水平明显下降,miRNA-204表达水平明显上升。结论 D-半乳糖诱导的衰老心肌细胞自噬水平降低的过程中,存在miRNA-30a、miRNA-126表达水平的降低以及miRNA-204表达水平的增加,miRNA的差异表达可能是衰老心肌细胞自噬水平变化的重要机制。
[Abstract]:Objective to investigate the relationship between microRNA (miRNA) and autophagy of aging cardiac myocytes. Methods the aging of rat cardiomyocytes was induced by 20 g / L D-galactose, 尾 -galactosidase staining was used to detect the aging level of cardiac myocytes, and the autophagy formation of cardiac myocytes induced by D-galactose was observed by electron microscope. The expression levels of miRNA-30asimiRNA-126 simiRNA-204 and autophagy LC3B / II- 3LCBI in aged rat cardiomyocytes and normal H9c2 cells were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blotting (Western blot), respectively. Results compared with the control group, cell staining deepened with the increase of drug concentration and stimulation days. When the concentration of D-galactose was 8 g / L for 9 days, the cell staining was the most obvious. The results of electron microscope showed that D-galactose induced decrease of autophagy in aging cardiomyocytes. Western blot showed that the level of LC3B II- / LCB I was decreased, and real-time fluorescence quantitative polymerase chain reaction showed that the level of LC3B 鈪，

本文编号：2085712