Ox-LDL对THP-1巨噬细胞迁移功能、microRNA21表达及MAPK通道磷酸化的影响  

Influence of ox-LDL on migration function of THP-1 macrophages and microRNA21 expression and MAPK pathway phosphorylation

作　　者：曾华甦 蔺艳军 高霖[1] 张田田[1] 曹嘉添 范虞琪[1] 殷兆芳[1] 顾俊[1] 王长谦[1] ZENG Hua-su, LIN Yan-jun, GAO Lin, ZHANG Tian-tian, CAO Jia-tian, FAN Yu-qi, YIN Zhao-fang, Gu Jun, WANG Chang-qian(Department of Cardiology, Affiliated Ninth Peoplels Hospital, Medical College of Shanghai Jiaotong University, Shanghai, 200011, China)

机构地区：[1]上海交通大学医学院附属第九人民医院心内科,上海200011

出　　处：《心血管康复医学杂志》2018年第3期241-246,共6页Chinese Journal of Cardiovascular Rehabilitation Medicine

基　　金：上海市科委重点项目(14JC1404400);高等学校博士学科点专项科研基金(20130073110016)~~

摘　　要：目的:探讨氧化型低密度脂蛋白(ox-LDL)对THP-1巨噬细胞迁移功能、microRNA21(miR-21)表达及丝裂原活化蛋白激酶(MAPK)通路的影响。方法:以160nmol/L佛波酯(PMA)诱导THP-1细胞分化成巨噬细胞。根据是否采用ox-LDL处理及脂质体介导miR-21抑制剂转染THP-1巨噬细胞,分为空白对照组(未进行oxLDL处理及脂质体转染),ox-LDL组(采用50mg/L ox-LDL处理,未进行脂质体转染),miR-21抑制剂组(脂质体介导miR-21抑制剂转染后采用50mg/L ox-LDL处理),miR-21抑制剂阴性对照组(脂质体介导miR-21抑制剂阴性对照转染后采用50mg/L ox-LDL处理)。Transwell法检测THP-1巨噬细胞迁移数,实时定量PCR检测miR-21表达情况,Western-blot法检测双特异性磷酸-8(DUSP-8)表达水平及磷酸化水平。结果:与空白对照组比较,ox-LDL组THP-1巨噬细胞迁移数显著增加[(74.10±15.10)比(184.10±26.28)],miR-21表达水平[(1.00±0.21)比(2.02±0.27)],MAPK通路的JNK、P38蛋白磷酸化水平显著升高,DUSP-8蛋白表达水平降低,P均<0.01;与ox-LDL组比较,miR-21抑制剂组THP-1巨噬细胞迁移数[(184.10±26.28)比(58.50±10.24)]显著减少,miR-21表达水平[(2.02±0.27)比(0.66±0.16)],JNK、P38蛋白磷酸化水平显著降低,DUSP-8蛋白表达水平显著升高,P均<0.01。结论:ox-LDL增强巨噬细胞迁移能力,可能与其上调miR-21表达、增强MAPK通路的JNK、P38蛋白磷酸化、降低DUSP-8表达有关。Objective：To explore influence of oxidized low density lipoprotein（ox-LDL）on migration function of THP-1 macrophages,expression of microRNA 21（miR-21）and mitogen-activated protein kinase（MAPK）pathway.Methods：Phorbol myristate acetate（PMA）of 160 nmol/L was used to induce THP-1 cells to differentiate into macrophages.According to application of ox-LDL treatment and liposomes-mediated miR-21 inhibitor transfecting THP-1 macrophages（transfection for short）or not,THP-1 macrophages were divided into blank control group（received neither ox-LDL treatment nor transfection）,ox-LDL group（received 50 mg/L ox-LDL treatment without transfection）,miR-21 inhibitor group（received 50 mg/L ox-LDL treatment after transfection）and miR-21 inhibitor negative-control group（received 50 mg/L ox-LDL treatment after negative-control transfection）.THP-1 macrophage migration number was measured by transwell method,miR-21 expression was measured by real-time quantitative PCR,and expression of dual specific phosphate 8（DUSP-8）and phosphorylation level of MAPK pathway were measured by Western-blot method.Results：Compared with blank control group,there were significant rise in migration number of THP-1 macrophages[（74.10±15.10）vs.（184.10±26.28）],miR-21 expression[（1.00±0.21）vs.（2.02±0.27）]and phosphorylation levels of JNK and P38 protein,and significant reduction in expression of DUSP-8 protein in ox-LDL group,P〈0.01 all.Compared with ox-LDL group,there were significant reductions in migration number of THP-1 macrophages[（184.10±26.28）vs.（58.50±10.24）],miR-21 expression [（2.02±0.27）vs.（0.66±0.16）]and phosphorylation levels of JNK and P38 protein,and significant rise in expression of DUSP-8 protein in ox-LDL group,P〈0.01 all.Conclusion：Ox-LDL enhances migration function of macrophages,which may be related to its effects of upregulating miR-21 expression,enhancing phosphorylation of JNK and P38 protein of MAPK pathway and reducing DUSP-8 expression.

关 键 词：脂蛋白类 LDL 巨噬细胞 微RNAS 

分 类 号：R341.350.9[医药卫生—基础医学]