Protective Effect and Mechanism of miR-328-3p on Coronary Artery Endothelial Cell Injury Induced by Oxidized Low-density Lipoprotein
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.
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维普
