沉默MAP4K4通过调控PPARγ/ABCA1通路缓解ox-LDL诱导的血管内皮细胞损伤  

Silencing of MAP4K4 alleviated ox-LDL-induced vascular endothelial cell injury via regulating PPARγ/ABCA1 signaling

作　　者：王玉香[1] 燕燕[1] 李永芳[1] 李艾帆[1] 李彦玲[1] WANG Yuxiang;YAN Yan;LI Yongfang;LI Aifan;LI Yanling(Department of Neurology,the First People’s Hospital of Zhengzhou,Zhengzhou,Henan 450004,China)

机构地区：[1]郑州市第一人民医院神经内科,河南省郑州市450004

出　　处：《中国动脉硬化杂志》2021年第1期54-59,共6页Chinese Journal of Arteriosclerosis

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20190983)。

摘　　要：目的探讨丝裂原活化蛋白激酶(MAP4K4)在氧化型低密度脂蛋白(ox-LDL)诱导的血管内皮细胞损伤中的作用及其机制。方法采用100 mg/L ox-LDL诱导人脐静脉内皮细胞(HUVEC)建立细胞损伤模型。RT-PCR检测HUVEC中MAP4K4的mRNA表达水平。Western blot法测定MAP4K4、Bax、Bcl-2、Cleaved Caspase-3(C-Caspase-3)、过氧化物酶体增殖物激活受体γ(PPARγ)和三磷酸腺苷结合盒转运体A1(ABCA1)蛋白表达量。CCK8法检测细胞存活率。ELISA试剂盒检测细胞凋亡。DCFH-DA用于检测细胞活性氧(ROS)水平。丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性分别用其试剂盒进行检测。结果ox-LDL能明显上调MAP4K4的表达。另外,沉默MAP4K4能够显著增加细胞活性,且抑制ox-LDL诱导的细胞凋亡;同时可抑制Bax和C-Caspase-3的蛋白表达并促进Bcl-2的蛋白表达量。此外,下调MAP4K4能够明显抑制ROS生成及MDA含量,并增加抗氧化酶SOD和CAT活性。机制研究显示沉默MAP4K4能够激活PPARγ/ABCA1信号通路。结论沉默MAP4K4通过激活PPARγ/ABCA1信号通路抑制ox-LDL诱导的HUVEC细胞凋亡和氧化应激从而减轻血管内皮细胞损伤,为治疗As提供理论依据。Aim The purpose of the present study was to investigate the potential role and mechanism of mitogen activated protein kinase kinase kinase kinase 4(MAP4K4)in human umbilical vein endothelial cell(HUVEC)injury induced by oxidized low density lipoprotein(ox-LDL).Methods The cell injury model was induced by ox-LDL(100 mg/L).The mRNA expression of MAP4K4 was detected using RT-PCR assay.Western blot analysis was conducted to estimate the protein levels of MAP4K4,Bax,Bcl-2,C-Caspase-3,peroxisome proliferators-activated receptorγ(PPARγ)and ATP binding cassette transporter A1(ABCA1).CCK8 assay was performed to measure cell viability.ELISA assay was applied to analyse cell apoptosis.The reactive oxygen species(ROS)production was measured by DCFH-DA assay.The content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)and catalase(CAT)were detected by commercially available assay kits.Results The expression of MAP4K4 was elevated in ox-LDL-stimulated HUVEC.In addition,silencing of MAP4K4 apparently enhanced cell viability and attenuated HUVEC apoptosis induced by ox-LDL accompanied by a decrease in the expression of Bax and C-Caspase-3 and an increase in the expression of Bcl-2.Moreover,MAP4K4 knockdown remarkably alleviated the generation of ROS and the content of MDA accompanied with the enhanced activity of anti-oxidative enzyme system SOD and CAT triggerd by ox-LDL.Mechanically,ablation of MAP4K4 obviously activated PPARγ/ABCA1 signaling pathway.Conclusion Depletion of MAP4K4 relieves ox-LDL-induced vascular endothelial injury by reducing apoptosis and mitigating oxidative damage via PPARγ/ABCA1 activation,thus providing the basis for the therapeutic effect of MAP4K4 on the As.

关 键 词：动脉粥样硬化 内皮细胞 MAP4K4 氧化应激 凋亡 

分 类 号：R541[医药卫生—心血管疾病]