Protective Effect and Mechanism of miR-328-3p on Coronary Artery Endothelial Cell Injury Induced by Oxidized Low-density Lipoprotein
- VernacularTitle:miR-328-3p对氧化型低密度脂蛋白诱导的冠状动脉内皮细胞损伤的保护作用及机制研究
- Author:
Yonglan HOU
1
;
Xia LI
;
Jianmei WANG
;
Zhen LIU
;
Minglei HAN
;
Zhenghao LIU
;
Weidong JIN
Author Information
- Keywords: miR-328-3p; Insulin-like growth factor 2; Oxidized low-density lipoprotein; Coronary artery endothelial cells
- From: Journal of Sichuan University (Medical Sciences) 2024;55(5):1210-1216
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the protective effect of miR-328-3p on oxidized low-density lipoprotein(ox-LDL)-induced coronary artery endothelial cell injury and the potentially relevant mechanisms.Methods Human coronary artery endothelial cells(HCAECs)were induced with ox-LDL,and the cells were divided into a control group consisting of normal cells,an ox-LDL group receiving ox-LDL treatment,an ox-LDL+miR-NC group transfected with miR-NC and treated with ox-LDL,an ox-LDL+miR-328-3p group transfected with miR-328-3p and treated with ox-LDL,and ox-LDL+miR-328-3p+pcDNA group co-transfected miR-328-3p and pcDNA and treated with ox-LDL,and an ox-LDL+miR-328-3p+insulin-like growth factor 2(IGF2)group co-transfected miR-328-3p and IGF2 and treated with ox-LDL.The expression level of miR-328-3p was determined with RT-qPCR.Cell proliferation was determined by MTT.Cell apoptosis was measured by flow cytometry.Western blot was conducted to examine the protein expression levels of cleaved cas-3 and IGF2.ELISA was performed to determine the levels of tumor necrosis factor α(TNF-α),interleukin(IL)-6,and IL-1β.Dual luciferase reporter experiment was performed to verify the targeting relationship between miR-328-3p and IGF2.Results Compared with those of the control group,miR-328-3p expression level and cell activity were significantly reduced in the ox-LDL group(P＜0.05),while the apoptotic rate,the protein expression levels of cleaved cas-3,IGF2,Bax,and Bcl-2,and the levels of TNF-α,IL-6,and IL-1β were significantly increased(P＜0.05).Compared with those of the ox-LDL+miR-NC group,miR-328-3p expression level and cell activity significantly increased in the ox-LDL+miR-328-3p group(P＜0.05),while the apoptosis rate,the protein expression levels of cleaved cas-3 and IGF2,and the levels of TNF-α,IL-6,and IL-1β were significantly reduced.IGF2 was a functional target of miR-328-3p.Compared with those of the ox-LDL+miR-328-3p+pcDNA co-transfection group,the IGF2 protein level was significantly increased(P＜0.05)and cell activity was significantly decreased(P＜0.05)in the ox-LDL+miR-328-3p+IGF2 co-transfection group,while the apoptosis rate,cleaved cas-3 protein level,and the levels of TNF-α,IL-6,and IL-1β were significantly elevated(P＜0.05).Conclusion miR-328-3p inhibits ox-LDL-induced apoptosis and inflammatory in coronary artery endothelial cell injury through targeted negative regulation of IGF2.
