过氧化物还原酶2通过ROS-NF-κB-miR-33a-ABCA1途径抑制巨噬细胞脂质蓄积  

Prdx2 Inhibits Macrophage Lipid Accumulation Through ROS-NF-κB-miR-33a-ABCA1 Pathway

作　　者：黄江伟 江鑫 蒋长荣[1] 龚慧琴[1] HUANG Jiang-Wei;JIANG Xin;JIANG Chang-Rong;GONG Hui-Qin(Department of Cardiovascularology,Affiliated Nanhua Hospital,University of South China,Hengyang 421002,Hunan,China;Department of Emergency,Affiliated Nanhua Hospital,University of South China,Hengyang 421002,Hunan,China)

机构地区：[1]南华大学附属南华医院心血管内科,湖南衡阳421002 [2]南华大学附属南华医院急诊科,湖南衡阳421002

出　　处：《中国生物化学与分子生物学报》2020年第12期1446-1454,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：衡阳市科技计划项目(No.2019jh010920)资助。

摘　　要：已有研究证实,过氧化物还原酶2(peroxiredoxin 2,Prdx2)可抑制小鼠动脉粥样硬化发展,但其在巨噬细胞脂质蓄积中的作用及机制未知。本文在体外培养THP-1源性泡沫细胞,转染Prdx2质粒过表达载体(pcDNA3.1-Prdx2)或Prdx2 siRNA,通过qRT-PCR、Western印迹、油红O和高效液相色谱等手段,检测ABCA1、NF-κB p65和miR-33a表达,胆固醇流出水平,胞内脂滴数目,胆固醇含量及ROS水平。此外,用NF-κB抑制剂PDTC和/或miR-33a抑制剂作预处理,观察上述指标有何变化。结果表明,Prdx2过表达组细胞ABCA1表达水平显著提高(P<0.05),而NF-κB和miR-33a水平明显下调(P<0.05),胞内[3H]-胆固醇流出增加(P<0.05),脂质蓄积减轻;而预处理PDTC和miR-33a抑制剂后,上述效应则更加明显。相反,Prdx2沉默组ABCA1水平下降(P<0.05),NF-κB和miR-33a表达上调(P<0.05)。我们发现,Prdx2过表达能有效降低泡沫细胞内ROS水平。综上所述,Prdx2可通过ROS-NF-κB-miR-33a-ABCA1途径促进巨噬细胞胆固醇流出,抑制脂质蓄积。Previous studies have unraveled that peroxiredoxin 2(Prdx2)inhibits atherogenesis in mice,whereas its role in macrophage lipid accumulation or the underlying mechanisms remain unknown.THP-1 monocyte-derived foam cells were transfected with Prdx2-overexpressing plasmid vectors(pcDNA3.1-Prdx2)or Prdx2 siRNA.The expression of ABCA1,NF-κB p65 and miR-33a were detected by RT-PCR and Western blotting.Percentage of cholesterol efflux was evaluated by liquid scintillation counting.Cellular lipid droplets were assessed using Oil Red O staining.Intracellular cholesterol contents were measured using high performance liquid chromatography(HPLC).Furthermore,cells were pre-treated with NF-κB inhibitor PDTC and/or miR-33a inhibitor,followed by detection of the indices above.The results showed that overexpression of Prdx2 in THP-1 monocyte-derived foam cells significantly increased ABCA1 expression and the percentage of[3H]-cholesterol efflux to apoA-1(P<0.05),whereas NF-κB p65 and miR-33a levels as well as lipid accumulation were decreased(P<0.05).After pre-treatment with PDTC and/or miR-33a inhibitor,these effects were more obvious(P<0.05).In contrast,silencing of Prdx2 significantly diminished ABCA1 expression and increased NF-κB p65 and miR-33a levels.At last,we found that Prdx2 overexpression obviously down-regulated the ROS level in THP-1 monocyte-derived foam cells.Altogether,Prdx2 promotes macrophage cholesterol efflux and inhibits intracellular lipid accumulation through the ROS-NF-κB-miR-33a-ABCA1 pathway.

关 键 词：过氧化物还原酶2 三磷酸腺苷结合盒转运体A1 活性氧 NF-ΚB miR-33a 

分 类 号：R363[医药卫生—病理学] R5[医药卫生—基础医学]