miR-193a-5p通过HDAC9-ABCA1/G1途径抑制巨噬细胞脂质蓄积  

miR-193a-5p Inhibits Macrophage Lipid Accumulation via the HDAC9-ABCA1/G1 Pathway

作　　者：王艳 宋志勇 艾卫敏 蔡金杏 WANG Yan;SONG Zhi-Yong;AI Wei-Min;CAI Jin-Xing(Department of Medical Basis,XiangTan Medicine&Health Vocational College,XiangTan 421002,Hunan,China)

机构地区：[1]湘潭医卫职业技术学院,医学基础课部,湖南湘潭411102

出　　处：《中国生物化学与分子生物学报》2024年第12期1751-1760,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：湖南省卫健委科研课题基金(No.202201024971)资助。

摘　　要：前人研究发现,微RNA-193a-5p(miR-193a-5p)参与上皮细胞脂肪酸代谢,但在巨噬细胞脂质蓄积中的作用未明。组蛋白去乙酰化酶9(HDAC9)被证实可通过对三磷酸腺苷结合盒转运子A1/G1(ABCA1/G1)进行去乙酰化修饰,抑制ABCA1/G1表达和巨噬细胞胆固醇外流,促进泡沫细胞形成。本研究探讨miR-193a-5p对THP-1巨噬细胞脂质蓄积的影响及其与HDAC9的联系。RT-PCR和Western印迹显示,THP-1源性泡沫细胞中miR-193a-5p水平显著下调,转染miR-193a-5p mimic可显著增加ABCA1和ABCG1的mRNA和蛋白质表达,而SR-BI、CD36和SR-A的表达无影响。油红O染色和液体闪烁计数法结果表明,过表达miR-193a-5p可显著减少胞内脂滴含量,而DiL-ox-LDL摄取水平无明显变化。高效液相色谱(HPLC)提示,miR-193a-5p mimic转染后泡沫细胞内总胆固醇(TC)、胆固醇酯(CE)和游离胆固醇(FC)含量均显著减少,上述效应在转染HDAC9 siRNA后显著增强(P<0.05),而在HDAC9过表达后被显著抵消(P<0.05)。荧光素酶报告基因检测和Western印迹提示,miR-193a-5p可通过靶向结合HDAC93′UTR,抑制其蛋白质表达(P<0.05)。综上,miR-193a-5p可通过HDAC9-ABCA1/G1途径抑制THP-1巨噬细胞脂质蓄积。Previous studies have shown that microRNA-193a-5p(miR-193a-5p)was related to fatty acid metabolism in epithelial cells,whereas its role in macrophage lipid accumulation is unclear.Histone deacetylase 9(HDAC9)could deacetylate ATP-binding cassette transporter A1/G1(ABCA1/G1)and decrease their expression,inhibiting macrophage cholesterol efflux and promoting foam cell formation.This study explores the effects of miR-193a-5p on lipid accumulation in THP-1 macrophages and its relationship with HDAC9.RT-PCR showed that miR-193a-5p levels were significantly decreased in THP-1-derived foam cells.RT-PCR and Western blotting results suggested that miR-193a-5p overexpression significantly up-regulated the mRNA and protein levels of ABCA1 and ABCG1,while it did not affect the expression of SR-A,SR-BI or CD36.Oil Red O staining and liquid scintillation counting indicated that miR-193a-5p mimic transfection markedly promoted cholesterol efflux and decreased lipid droplet numbers without altering the uptake of DiL-ox-LDL.High-performance liquid chromatography(HPLC)assay showed that miR-193a-5p overexpression decreased intracellular contents of total cholesterol(TC),cholesterol ester(CE),and free cholesterol(FC)in THP-1-derived foam cells.These inhibitory effects of miR-193a-5p on foam cell formation became more evident after HDAC9 siRNA treatment and were mainly reversed by HDAC9 overexpression(P<0.05).Furthermore,luciferase reporter assay and Western blotting showed that miR-193a-5p directly targeted HDAC93′UTR,thereby down-regulating its protein expression(P<0.05).Altogether,miR-193a-5p inhibits lipid accumulation in THP-1 macrophages via the HDAC9-ABCA1/G1 pathway.

关 键 词：miR-193a-5p 组蛋白去乙酰化酶9 三磷酸腺苷结合盒转运子A1/G1 脂质蓄积 巨噬细胞 

分 类 号：Q591.5[生物学—生物化学]