lncRNA LINC00339通过下调miR-149-5p表达调控ox-LDL诱导的人脑微血管内皮细胞损伤  

lncRNA LINC00339 regulates ox-LDL-induced human brain microvas⁃cular endothelial cell damage by down-regulating miR-149-5p expression

作　　者：王进鹏[1] 龙启福 雷延成 张豪[1] 林存山[1] WANG Jin-peng;LONG Qi-fu;LEI Yan-cheng;ZHANG Hao;LIN Cun-shan(Department of Neurology,Qinghai People's Hospital,Qinghai Uni-versity,Xining 810007,China;Basic Medical Research Center of Medical College,Qinghai Uni-versity,Xining 810007,China)

机构地区：[1]青海省人民医院神经内科,青海西宁810007 [2]青海大学医学院基础医学研究中心,青海西宁810007

出　　处：《中国病理生理杂志》2021年第2期277-283,共7页Chinese Journal of Pathophysiology

基　　金：青海省科技计划项目(No.2014-NS-120-1)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)LINC00339对氧化型低密度脂蛋白(ox-LDL)诱导的人脑微血管内皮细胞(HBMEC)损伤的影响及分子机制。方法:将HBMEC分为对照组(未经任何处理)、ox-LDL(50 mg/L)组、ox-LDL+si-NC组(转染si-NC+50 mg/L ox-LDL)、ox-LDL+si-LINC00339组(转染si-LINC00339+50 mg/L ox-LDL)、ox-LDL+miR-NC组(转染miR-NC+50 mg/L ox-LDL)、ox-LDL+miR-149-5p组(转染miR-149-5p+50 mg/L ox-LDL)、ox-LDL+si-LINC00339+anti-miR-NC组(共转染si-LINC00339和anti-miR-NC+50 mg/L ox-LDL)及ox-LDL+si-LINC00339+anti-miR-149-5p组(共转染si-LINC00339和anti-miR-149-5p+50 mg/mL ox-LDL)。RT-qPCR检测LINC00339和miR-149-5p的表达水平;分别检测丙二醛(MDA)含量及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;流式细胞术检测细胞凋亡;Western blot法检测Bcl-2和Bax蛋白表达;双萤光素酶报告基因实验检测LINC00339和miR-149-5p的靶向关系。结果:ox-LDL诱导的HBMEC中LINC00339的表达水平升高,miR-149-5p的表达水平降低(P<0.05)。抑制LINC00339表达或过表达miR-149-5p可致HBMEC的MDA含量降低,SOD和CAT活性升高,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低(P<0.05)。LINC00339靶向调控miR-149-5p,敲减miR-149-5p表达逆转了抑制LINC00339表达对ox-LDL诱导的HBMEC氧化损伤的作用。结论:抑制LINC00339表达可能通过上调miR-149-5p表达抑制ox-LDL诱导的HBMEC凋亡和氧化应激反应,从而减轻细胞损伤。AIM:To investigate the effect of long noncoding RNA(lncRNA)LINC00339 on human brain microvascular endothelial cell(HBMEC)damage induced by oxidized low-density lipoprotein(ox-LDL)and its molecular mechanism.METHODS:The HBMEC was divided into control group,ox-LDL group,si-NC group,si-LINC00339 group,pcDNA group,pcDNA-LINC00339 group,ox-LDL+si-NC group,ox-LDL+si-LINC00339 group,ox-LDL+miR-NC group,ox-LDL+miR-149-5p group,ox-LDL+si-LINC00339+anti-miR-NC group,and ox-LDL+si-LINC00339+anti-miR-149-5p group.The expression of LINC00339 and miR-149-5p was detected by RT-qPCR.The content of malonaldehyde(MDA),and the activity of superoxide dismutase(SOD)and catalase(CAT)were measured by kits.The apoptosis was analyzed by flow cytometry.The protein expression of Bcl-2 and Bax was determined by Western blot.Dual-luciferase reporter experiment was used to detect the targeting relationship between LINC00339 and miR-149-5p.RESULTS:The expression of LINC00339 in HBMEC induced by ox-LDL was increased,while the expression of miR-149-5p was decreased(P<0.05).Inhibition of LINC00339 expression or miR-149-5p over-expression reduced MDA content,increased SOD and CAT activity,decreased apoptotic rate,increased Bcl-2 expression and decreased Bax expression(P<0.05).LINC00339 targeted miR-149-5p and regulated miR-149-5p expression.Knock-down of miR-149-5p expression reversed the effect of LINC00339 expression inhibition on ox-LDL-induced oxidative damage of HBMEC.CONCLUSION:Inhibition of LINC00339 expression may inhibit ox-LDL-induced apoptosis and oxidative stress in HBMEC by up-regulating the expression of miR-149-5p,which may have a protective effect on HBMEC.

关 键 词：LINC00339 微小RNA-149-5p 人脑微血管内皮细胞 氧化应激 细胞凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学] R363[医药卫生—临床医学]