Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, No. 110 Xiang-Ya Road, Changsha 410078, China.
Alcohol. 2013 Sep;47(6):481-93. doi: 10.1016/j.alcohol.2013.05.005. Epub 2013 Jul 3.
MicroRNAs (miRNAs) have been shown to be closely associated with cellular apoptosis, but their involvement in response to ethanol-induced gastric mucosal epithelial cell apoptosis remains largely unknown. The purpose of this study was to investigate the expression profile of apoptosis-associated miRNAs in ethanol-induced acute gastric mucosal injury and the mechanisms underlying injury. Gastric mucosal injury was induced in rats by oral administration of ethanol, and gastric tissues were collected for analysis of gastric ulcer index, apoptosis ratio, caspase-3 activity, and miRNAs expression. Cell cultures of human gastric mucosal epithelial cells (GES-1) were incubated with ethanol to induce apoptosis. Mimics or inhibitors of miRNAs or c-Jun N-terminal kinase (JNK) inhibitor were added to the cell culture medium. GES-1 cells were collected for analysis of apoptosis ratio, caspase-3 activity, miRNAs expression, and protein phosphorylation levels of JNK, p38 mitogen-activated protein kinase (p38MAPK), or extracellular signal-regulated kinase (ERK). In the animal experiments, gastric ulcer index, cellular apoptosis, and caspase-3 activity were significantly increased, accompanied by up-regulation of miR-145 and down-regulation of the microRNAs miR-17, miR-19a, miR-21, miR-181a, and miR-200c. In the human cell culture experiments, the anti-apoptotic effects of miR-19a and miR-21 or pro-apoptotic effect of miR-145 were confirmed by their corresponding mimics or inhibitor; the ethanol-induced GES-1 apoptosis as well as the changes in miRNAs expression were significantly attenuated in the presence of JNK inhibitor. These results demonstrated that miR-145, miR-19a, and miR-21 were the apoptosis-associated miRNAs in gastric mucosal epithelial cells. The regulation of expression of these 3 miRNAs in ethanol-induced GES-1 apoptosis involved the JNK pathway.
微小 RNA(miRNAs)与细胞凋亡密切相关,但它们在乙醇诱导的胃黏膜上皮细胞凋亡反应中的作用仍知之甚少。本研究旨在探讨与细胞凋亡相关的 miRNAs 在乙醇诱导的急性胃黏膜损伤及其损伤机制中的表达谱。通过口服乙醇诱导大鼠胃黏膜损伤,收集胃组织分析胃黏膜损伤指数、细胞凋亡率、caspase-3 活性和 miRNAs 表达。将人胃黏膜上皮细胞(GES-1)进行乙醇孵育以诱导细胞凋亡,加入 miRNAs 模拟物或抑制剂或 c-Jun N-末端激酶(JNK)抑制剂至细胞培养液中。收集 GES-1 细胞分析细胞凋亡率、caspase-3 活性、miRNAs 表达以及 JNK、p38 丝裂原激活蛋白激酶(p38MAPK)或细胞外信号调节激酶(ERK)的磷酸化水平。在动物实验中,胃黏膜损伤指数、细胞凋亡和 caspase-3 活性显著增加,同时 miR-145 上调,miR-17、miR-19a、miR-21、miR-181a 和 miR-200c 下调。在人细胞培养实验中,通过相应的模拟物或抑制剂证实了 miR-19a 和 miR-21 的抗凋亡作用和 miR-145 的促凋亡作用;JNK 抑制剂存在时,乙醇诱导的 GES-1 凋亡以及 miRNAs 表达的变化明显减弱。这些结果表明,miR-145、miR-19a 和 miR-21 是胃黏膜上皮细胞中的凋亡相关 miRNAs。这些 miRNAs 在乙醇诱导的 GES-1 细胞凋亡中的表达调控涉及 JNK 途径。