本文選題：衰老心肌細(xì)胞 + 自噬��； 參考：《中國(guó)動(dòng)脈硬化雜志》2017年04期

【摘要】：目的探討microRNA(miRNA)和衰老心肌細(xì)胞自噬之間的關(guān)系。方法使用0、4、8、16、20 g/L D-半乳糖刺激大鼠心肌細(xì)胞H9c2衰老,刺激3、6、9天后,β-半乳糖苷酶染色法檢測(cè)細(xì)胞衰老水平;電鏡觀察D-半乳糖刺激衰老大鼠心肌細(xì)胞自噬體形成;分別采用實(shí)時(shí)熒光定量PCR(qRT-PCR)及蛋白質(zhì)印跡法(Western blot)檢測(cè)衰老大鼠心肌細(xì)胞與正常H9c2細(xì)胞中miRNA-30a、miRNA-126、miRNA-204及自噬蛋白LC3B II/LC3B I表達(dá)水平。結(jié)果與對(duì)照組比較,隨著藥物濃度的升高及刺激天數(shù)的增加,細(xì)胞染色逐漸加深,在D-半乳糖濃度為8 g/L刺激9天時(shí),細(xì)胞染色最明顯;電鏡結(jié)果顯示,D-半乳糖誘導(dǎo)衰老心肌細(xì)胞自噬體減少;Western blot結(jié)果顯示,衰老心肌細(xì)胞LC3B II/LC3B I水平降低;實(shí)時(shí)熒光定量PCR結(jié)果顯示,誘導(dǎo)衰老組心肌細(xì)胞中,miRNA-30a、miRNA-126表達(dá)水平明顯下降,miRNA-204表達(dá)水平明顯上升。結(jié)論 D-半乳糖誘導(dǎo)的衰老心肌細(xì)胞自噬水平降低的過(guò)程中,存在miRNA-30a、miRNA-126表達(dá)水平的降低以及miRNA-204表達(dá)水平的增加,miRNA的差異表達(dá)可能是衰老心肌細(xì)胞自噬水平變化的重要機(jī)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，

本文編號(hào)：2085712