Yuan Linhui, Yu Leitao, Zhang Jing, Zhou Zhidong, Li Chang, Zhou Bin, Hu Xiaolan, Xu Guohai, Tang Yanhua
Department of Anesthesiology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Department of Thyroid Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2020 Apr;21(4):1709-1716. doi: 10.3892/mmr.2020.10978. Epub 2020 Feb 6.
Myocardial ischemia/reperfusion injury often leads to adverse cardiovascular outcomes due to severe hypoxia. The present study aimed to evaluate the effects and mechanism of long non‑coding RNA H19 (H19) on rat H9c2 cells with hypoxia‑induced injury. H9c2 cells were infected with lentiviruses to express H19 or H19‑targeting short hairpin RNA (shRNA), or their respective controls, at a multiplicity of infection of 1:100. H19 expression was determined by reverse transcription‑quantitative PCR. Hypoxic injury was induced and assessed by analyzing the level of apoptosis, the cell cycle distribution and the mitochondrial membrane potential using flow cytometry in the different groups. The expression of the PI3K/AKT and the ERK/p38 signaling pathways were analyzed using western blotting. It was found that hypoxia stimulated apoptosis, induced G1 phase cell cycle arrest and increased the mitochondrial depolarization rate in H9c2 cells. When compared with the hypoxic model group, the H19 overexpression group had a significantly reduced rate of apoptosis (P=0.016), a smaller G1 population and a higher S phase population (P=0.018 and P=0.031, respectively), and a reduced mitochondrial depolarization rate (P=0.036). By contrast, the H19 shRNA group exhibited the opposite trends, suggesting that hypoxia‑induced injury was alleviated by the overexpression of H19 and was aggravated by the knockdown of H19. The present mechanistic studies revealed that H19 may decrease hypoxia‑induced cell injury by activating the PI3K/AKT and ERK/p38 pathways. The results of the present study suggested that H19 may alleviate hypoxia‑induced myocardial cell injury through the activation of the PI3K/AKT and ERK/p38 pathways.
心肌缺血/再灌注损伤常因严重缺氧导致不良心血管结局。本研究旨在评估长链非编码RNA H19（H19）对缺氧诱导损伤的大鼠H9c2细胞的影响及其机制。以1:100的感染复数用慢病毒感染H9c2细胞，使其表达H19或靶向H19的短发夹RNA（shRNA），或各自的对照。通过逆转录定量PCR测定H19表达。通过流式细胞术分析不同组细胞的凋亡水平、细胞周期分布和线粒体膜电位，诱导并评估缺氧损伤。使用蛋白质印迹法分析PI3K/AKT和ERK/p38信号通路的表达。结果发现，缺氧刺激H9c2细胞凋亡，诱导G1期细胞周期阻滞并增加线粒体去极化率。与缺氧模型组相比，H19过表达组的凋亡率显著降低（P = 0.016），G1期细胞群体较小，S期细胞群体较高（分别为P = 0.018和P = 0.031），线粒体去极化率降低（P = 0.036）。相比之下，H19 shRNA组呈现相反趋势，表明H19过表达可减轻缺氧诱导的损伤，而H19敲低则加重损伤。目前的机制研究表明，H19可能通过激活PI3K/AKT和ERK/p38通路来减少缺氧诱导的细胞损伤。本研究结果提示，H19可能通过激活PI3K/AKT和ERK/p38通路减轻缺氧诱导的心肌细胞损伤。
