作者：韩婕1 ，高登峰 1 ，王晓娜 2 ，曹瑞华 2 ，邓捷 1 ，叶平 2

单位：

1.710004 陕西省西安市，西安交通大学第二附属医院心血管内科；　2.100853 北京市，中国人民解放军总医院第二医学中心老年心血管内科；通信作者：韩婕，E-mail：15909205752@163.com

关键词：

脂代谢障碍；胆固醇外流；miRNA-638；巨噬细胞

Keywords：

Lipid metabolism disorders；Cholesterol efflux；miRNA-638；Macrophages

CLC：

R 589

DOI：

DOI：10.3969/j.issn.1008-5971.2020.05.013

Funds：

国家自然科学基金资助项目（81570382）；陕西省重点研发计划（2017SF-134）

摘要：

背景　流行病学研究表明生活方式、饮食结构等外部因素的改变与遗传因素相互作用导致人体脂代谢紊乱，而胆固醇逆转运过程能够维持体内胆固醇稳态。目的　探讨 miRNA-638 对巨噬细胞内胆固醇外流的影响及其发生机制。方法　实验于 2016 年 9 月—2017 年 9 月完成。采用 100 ng/ml 佛波酯诱导人髓系白血病单核细胞（THP-1）贴壁并分化为巨噬细胞，根据预实验结果选取浓度为 20 nmol/L 的 miRNA-638 模拟物及其阴性对照物进行转染。采用 NBD- 胆固醇分别检测转染 miRNA-638 模拟物及其阴性对照物巨噬细胞经载脂蛋白 AI（ApoAI）、高密度脂蛋白（HDL）及标准血清外流液介导的胆固醇外流率；采用实时荧光定量 PCR 及 Western blotting 检测转染 miRNA-638 模拟物及其阴性对照物巨噬细胞中三磷酸腺苷结合盒转运体 A1（ABCA1）、三磷酸腺苷结合盒转运体 G1（ABCG1）和极低密度脂蛋白受体（VLDLR）mRNA 及其相对表达量。结果　转染 miRNA-638 模拟物巨噬细胞经 ApoAI 外流液介导的 NBD- 胆固醇外流率低于转染阴性对照物巨噬细胞（P<0.05）；转染 miRNA-638 模拟物巨噬细胞与转染阴性对照物巨噬细胞经 HDL、标准血清外流液介导的 NBD- 胆固醇外流率比较，差异无统计学意义（P>0.05）。转染miRNA-638 模拟物巨噬细胞的 ABCA1、ABCG1、VLDLR mRNA 相对表达量低于转染阴性对照物巨噬细胞（P<0.05）。转染 miRNA-638 模拟物巨噬细胞中 ABCA1、ABCG1 相对表达量低于转染阴性对照物巨噬细胞（P<0.05）；转染miRNA-638 模拟物巨噬细胞与转染阴性对照物巨噬细胞中 VLDLR 相对表达量比较，差异无统计意义（P>0.05）。结论　miRNA-638可减少由ApoAI介导的巨噬细胞内胆固醇外流，该作用可能是通过调控ABCA1/ABCG1的表达实现的。

Abstract：

Background　Epidemiological studies have shown that the changed way of life and diet，which interactwith genetic factors，lead to increased lipid metabolism disorders in population.Whereas reverse cholesterol transport could helpto maintain cholesterol homeostasis.Objective　To investigate the impact of miRNA-638 on cholesterol efflux from macrophagesand its mechanism.Methods　This experiment was conducted from September 2016 to September 2017.THP-1 cell line wastreated with 100 ng/ml PMA to differentiate into macrophage and then transfected with 20 nmol/L miRNA-638 mimics or itsnegative control.NBD-cholesterol was used to detect the effects of miRNA-638 on intracellular cholesterol efflux rate inducedby three different medias including apolipoprotein AI （ApoAI），HDL and standard serum.Relative expressions of ABCA1，ABCG1 and VLDLR mRNA and protein in transfected miRNA-638 mimicry and negative control macrophages were detectedby fluorescence real-time quantitative PCR and Western blotting.Results　NBD-cholesterol efflux rate by ApoAI efflux intransfected miRNA-638 mimicry macrophages was statistically significantly lower than that in transfected negative controlmacrophages（P<0.05），but there was no statistically significant difference in NBD-cholesterol efflux rate by HDL，standardserum between transfected miRNA-638 mimics and negative control macrophages（P>0.05）.Relative expression levels ofABCA1，ABCG1 and VLDLR mRNA in transfected miRNA-638 mimics macrophages were statistically significantly lower thanthose in transfected negative control macrophages（P<0.05）.Protein expression levels of ABCA1 and ABCG1 in transfectedmiRNA-638 mimics macrophages were statistically significantly lower than those in transfected negative control macrophages（P<0.05），however，there was no statistically significant difference in protein expression level of VLDLR betweenmiRNA-638 mimics macrophages and negative control macrophages（P>0.05）.Conclusion　miRNA-638 reduces cholesterolefflux from macrophages mediated by ApoAI，which may be achieved by regulating the protein expression levels of ABCA1/ABCG1.

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