miR-195-5p通过靶向调控FBXW7基因对氧化型低密度脂蛋白诱导的血管内皮细胞损伤的影响研究  

Effects of miR-195-5p on ox-LDL-induced vascular endothelial cell injury through regulating FBXW7

作　　者：林娜萍 吴玉塘[1] 黄德奋[1] 许朝祥[1] LIN Naping;WU Yutang;HUANG Defen;XU Chaoxiang(Department of Cardiology,Fujian Medical University 2nd affiliated Hospital,Quanzhou 362000,Fujian,China)

机构地区：[1]福建医科大学附属第二医院心内科,福建泉州362000

出　　处：《中国临床药理学与治疗学》2022年第2期163-170,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：福建省自然科学基金项目(2020J01228)。

摘　　要：目的:探讨miR-195-5p对氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的影响及其作用机制。方法:RT-PCR检测miR-195-5p在ox-LDL诱导的HUVECs细胞中的表达;CCK-8、ELISA和流式细胞仪分别检测细胞增殖、氧化应激和凋亡;双荧光酶素报告分析实验检测miR-195-5p的潜在作用靶点;Western blot检测靶蛋白在ox-LDL诱导的HUVECs细胞中的表达及其与miR-195-5p表达之间的关系。结果:miR-195-5p在ox-LDL诱导的HUVECs细胞中的表达显著高于正常HUVECs细胞。随后,在ox-LDL诱导的HUVECs细胞中沉默miR-195-5p能有效促进HUVECs细胞增殖,并抑制氧化应激因子[丙二醛(malondialdehyde,MDA)和乳酸脱氢酶(lactate dehydrogenase,LDH)]表达和凋亡水平。双荧光酶素实验和Western blot结果分别显示miR-195-5p可以直接靶向FBXW7蛋白基因并负调控FBXW7的表达。最后,miR-195-5p能通过调节FBXW7的表达影响ox-LDL诱导的HUVECs细胞增殖、氧化应激和凋亡。结论:miR-195-5p在ox-LDL诱导的HUVECs细胞中高表达。此外,miR-195-5p能通过抑制FBXW7的表达来加剧ox-LDL诱导的HUVECs细胞毒性、氧化应激和凋亡损伤。AIM:To investigate the effect of miR-195-5p on ox-LDL-induced vascular endothelial cell injury and its mechanism.METHODS:The expression of miR-195-5p in HUVECs cells induced by ox-LDL was detected by RT-PCR.Cell proliferation,the expression of oxidative stress associated factors(MDA and LDH)and apoptosis were detected by CCK-8,ELISA and flow cytometry,respectively.The potential targets of miR-195-5p were determined by dual luciferase reporter assay.The expression of target protein in ox-LDL-induced HUVECs cells and the relationship between miR-195-5p and FBXW7 expression were detected by Western blotting.RESULTS:The expression of miR-195-5p in ox-LDL-induced HUVECs cells was significantly higher than that in normal HUVECs cells.Subsequently,miR-195-5p silencing in ox-LDL-induced HUVECs cells effectively promoted the proliferation of HUVECs cells,whereas suppressed the expression of oxidative stress associated factors(MDA and LDH)and apoptosis level.Luciferase reporter assay and Western blot results showed that miR-195-5p could directly target FBXW7 protein gene and negatively regulate its expression.Finally,miR-195-5p modulated the proliferation,oxidative stress and apoptosis of HUVECs cells induced by ox-LDL by regulating the expression of FBXW7.CONCLUSION:miR-195-5p is highly expressed in HUVECs cells induced by ox-LDL.In addition,miR-195-5p can aggravate ox-LDL-induced cytotoxicity,oxidative stress and apoptosis of HUVECs cells by inhibiting the expression of FBXW7.

关 键 词：miR-195-5p FBXW7 氧化型低密度脂蛋白 血管内皮细胞 氧化应激 

分 类 号：R965.2[医药卫生—药理学]