褪黑素通过PPARγ-LXRα调控THP-1巨噬细胞ABCA1表达  

Melatonin Regulates the Expression of the THP-1 Derived Macrophage ABCA1 Receptor through PPARγ-LXRα

作　　者：王雨 陈连凤 王伟 宋玮 严晓伟 Wang Yu;Chen Lianfeng;Wang Wei(Department of Cardiology,Fu Wai Hospital,National Center for Cardiovascular Disease,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,China)

机构地区：[1]中国医学科学院/北京协和医学院国家心血管病中心、中国医学科学院阜外医院心内科,100037 [2]中国医学科学院/北京协和医学院北京协和医院心内科,1000730

出　　处：《医学研究杂志》2021年第7期26-30,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(面上项目)(81270381)。

摘　　要：目的研究褪黑素对巨噬细胞ABCA1受体表达的影响及作用机制。方法使用THP-1细胞体外培养PMA诱导分化为巨噬细胞,给予不同浓度的褪黑素(0、50nmol/L、100nmol/L、500nmol/L和1μmol/L)干预细胞,RT-PCR和Western blot法分别检测巨噬细胞ABCA1和LXRα的mRNA和蛋白相对表达量的变化。分别应用LXRα抑制剂GSK2033、PPARγ抑制剂GW9662、非选择性MT1/MT2受体拮抗剂Luzindole、选择性MT2受体拮抗剂K185进一步验证褪黑素调控ABCA1表达的作用机制。结果褪黑素显著上调THP-1来源巨噬细胞ABCA1和LXRα的mRNA和蛋白相对表达量,且呈剂量依赖性(P<0.05)。GW9662均可显著抑制褪黑素上调的ABCA1和LXRα蛋白表达水平(P=0.000),GSK2033可显著抑制褪黑素上调的ABCA1蛋白表达(P=0.000),但对LXRα蛋白表达水平无影响。而Luzindole和K185均未明显抑制褪黑素上调的ABCA1蛋白表达。结论褪黑素显著上调巨噬细胞ABCA1的表达,PPARγ-LXRα通路参与了褪黑素对ABCA1表达的调节作用。Objective To study the effect of melatonin on macrophage ABCA1 receptor expression and its mechanism.Methods THP-1 cells were cultured in vitro and differentiated into macrophages by PMA.The cells were treated with melatonin at different concentrations(0,50nmol/L,100nmol/L,500nmol/L and 1μmol/L,respectively).The mRNA and protein levels of ABCA1 and LXRα were measured by RT-PCR and Western Blot analysis,respectively.LXRα inhibitor GSK2033,PPARγ inhibitor GW9662,non-selective MT1/MT2 receptor antagonist Luzindole and selective MT2 receptor antagonist K185 were used to further verify the mechanism of melatonin in regulating ABCA1 expression.Results Melatonin significantly upregulated the relative mRNA and protein expression levels of ABCA1 and LXRα of macrophages in a dose-dependent manner(P<0.05).GW9662 significantly inhibited the upregulated ABCA1 and LXRα protein levels(P=0.000),and GSK2033 significantly inhibited the upregulated ABCA1 and LXRα protein levels(P=0.000),but had no effect on LXRα protein expression levels.Neither Luzindole nor K185 significantly inhibited the expression of ABCA1 protein upregulated by melatonin.Conclusion Melatonin significantly upregulated the expression of ABCA1 in macrophages,and PPARγ-LXRα pathway was involved in the regulation of ABCA1 expression by melatonin.

关 键 词：褪黑素 ABCA1 PPARΓ LXRΑ 

分 类 号：R3[医药卫生—基础医学]