Zhu Zhongsheng, Li Jinyu, Tong Rui, Zhang Xiaorong, Yu Bo
Department of Cardiology, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Pudong New District, Shanghai 201399, China.
Department of Vascular Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Pudong New District, Shanghai 201399, China.
Cardiol Res Pract. 2021 Aug 26;2021:9963258. doi: 10.1155/2021/9963258. eCollection 2021.
Atherosclerosis is a chronic process that takes place in the vascular wall and causes various cardiovascular diseases (CVDs). Micro-RNA-149 (miR-149) mediates many physiological and pathological processes, including atherosclerosis. However, it is unclear about the roles of miR-149 in endothelial injury. Here, we explored the protective effect and related mechanism of miR-149 in endothelial cells induced with oxidized low-density lipoprotein (ox-LDL).
Human endothelial cell lines (HUVECs) were exposed to ox-LDL to induce endothelial injury. Cell viability was determined by the CCK-8 assay. Autophagy was detected by immunofluorescence. RT-qPCR and western blot were carried out to determine the mRNA and protein expressions of Akt and mTOR.
The miR-149 level in HUVECs was reduced by ox-LDL (100 g/mL) incubation in a time-dependent manner. miR-149-mimic transfection markedly protected HUVECs from ox-LDL-induced injury, with increased cell viability and reduced caspase-3 activity. miR-149 mimics enhanced HUVEC autophagy, which was induced initially by ox-LDL. miR-149 mimics also markedly downregulated the expression of Akt, p-Akt, mTOR, and p-mTOR in ox-LDL-treated HUVECs. The miR-149-induced protection against HUVECs injury could be reversed by cotreatment with 3-methyladenine (3-MA, an autophagy inhibitor) or insulin (an activator of Akt/mTOR pathway).
miR-149 prevents ox-LDL-induced endothelial cell injury by enhancing autophagy via increasing Akt and mTOR expressions.
动脉粥样硬化是一种发生在血管壁的慢性过程,可导致各种心血管疾病(CVD)。微小RNA-149(miR-149)介导许多生理和病理过程,包括动脉粥样硬化。然而,miR-149在内皮损伤中的作用尚不清楚。在此,我们探讨了miR-149在氧化型低密度脂蛋白(ox-LDL)诱导的内皮细胞中的保护作用及相关机制。
将人内皮细胞系(HUVECs)暴露于ox-LDL以诱导内皮损伤。通过CCK-8法测定细胞活力。通过免疫荧光检测自噬。进行RT-qPCR和蛋白质印迹以确定Akt和mTOR的mRNA和蛋白质表达。
ox-LDL(100μg/mL)孵育以时间依赖性方式降低了HUVECs中的miR-149水平。miR-149模拟物转染显著保护HUVECs免受ox-LDL诱导的损伤,细胞活力增加且caspase-3活性降低。miR-149模拟物增强了ox-LDL最初诱导的HUVEC自噬。miR-149模拟物还显著下调了ox-LDL处理的HUVECs中Akt、p-Akt、mTOR和p-mTOR的表达。与3-甲基腺嘌呤(3-MA,一种自噬抑制剂)或胰岛素(Akt/mTOR途径的激活剂)共同处理可逆转miR-149诱导的对HUVECs损伤的保护作用。
miR-149通过增加Akt和mTOR表达增强自噬来预防ox-LDL诱导的内皮细胞损伤。