分化拮抗非编码RNA-201对单核-巨噬细胞功能的调控研究  

Regulation of Macrophages Functions by DANCR-201

作　　者：周海清 刘洋[2] 陈宇[2] 李荣霞 杨淑均 张伟丽[2] ZHOU Haiqing;LIU Yang;CHEN Yu;LI Rongxia;YANG Shujun;ZHANG Weili(Medical College,Anhui University of Science and Technology,Huainan(232000),Anhui,China)

机构地区：[1]安徽理工大学医学院,安徽省淮南市232000 [2]北京协和医学院中国医学科学院国家心血管病中心阜外医院心血管疾病国家重点实验室 [3]厦门大学附属心血管病医院

出　　处：《中国循环杂志》2020年第7期677-684,共8页Chinese Circulation Journal

基　　金：国家自然科学基金(81670338;91339101)。

摘　　要：目的:探究分化拮抗非编码RNA-201(differentiation antagonizing non-protein coding RNA,DANCR-201)在单核-巨噬细胞炎症反应中的调控作用。方法:纳入中国医学科学院阜外医院疑为冠心病接受冠状动脉CT检查的患者25例,年龄≥45岁。根据冠状动脉CT血管造影图像,研究对象分为3组,包括无斑块健康对照组(n=10)、完全钙化斑块组(n=10)和易损斑块组(n=5)。采用全转录组测序技术分析外周血全血中表达差异的长链非编码RNA分子。培养人单核细胞白血病细胞系(THP)-1细胞,用于建立M1和M2型巨噬细胞极化模型,采用慢病毒感染和Smart Silencer转染技术分别过表达和敲低DANCR-201,检测炎症因子、脂质代谢与血管钙化相关基因的表达水平,采用油红O染色法检测巨噬细胞脂质吞噬能力,采用胆固醇流出实验检测巨噬细胞胆固醇逆转运能力。结果:DANCR-201在易损斑块组患者外周血的表达水平显著升高。在单核细胞系中过表达DANCR-201,可促进单核趋化蛋白-1(MCP-1)和肿瘤坏死因子-α(TNF-α)高表达(P<0.01);反之,敲低DANCR-201时,可使MCP-1和TNF-α的表达水平显著下降(P<0.01)。在M1和M2型巨噬细胞中,DANCR-201显著抑制Runt相关转录因子2(Runx 2)表达水平(P<0.01),并促进骨保护素(OPG)高表达(P<0.05);反之,敲低DANCR-201时,M1和M2型巨噬细胞中的Runx 2表达水平升高,而OPG表达水平下降(P<0.01)。DANCR-201对巨噬细胞脂质吞噬和胆固醇逆转运能力无显著影响。结论:DANCR-201促进M1和M2型巨噬细胞的炎症反应,其在动脉粥样硬化斑块形成中的作用机制仍需深入研究。Objectives:To determine whether the differentiation antagonizing non-protein coding RNA 201(DANCR-201)is involved in the process of atherosclerosis through regulating the inflammation and function of macrophages.Methods:The whole-transcriptional RNA sequencing was used to analyze the differential expression level of long noncoding RNAs in peripheral whole blood from individuals with vulnerable plaque,or calcified plaque,or without plaques in the coronary arteries.The human acute monocytic leukemia cell lines(THP-1)were cultured.The overexpression of DANCR-201 was conducted by transfecting with DANCR-201 recombinant lentiviruses and the knockdown conducted by transfecting with DANCR-201 smart silencer.The real-time PCR method was performed to detect the mRNA expression of cytokine genes related to inflammation,cholesterol homeostasis and vascular calcification.Lipid phagocytosis of macrophages was observed by the oil red O staining method,and cholesterol efflux experiment was performed to assess the reverse cholesterol transport ability of macrophages.Results:The expression level of DANCR-201 was significantly higher in peripheral whole blood from patients with coronary vulnerable plaques than that from patients with calcified plaques or healthy subjects without plaques.In vitro experiments of THP-1 cells showed that overexpressing DANCR-201 significantly increased the expression levels of monocyte chemotactic protein-1(MCP-1)and tumor necrosis factor alpha(TNF-α)(P<0.01),and conversely,knockdown of DANCR-201 significantly inhibited the expression of MCP-1 and TNF-α(P<0.01).In M1 and M2 macrophages,overexpressing DANCR-201 inhibited the expression of runt-related transcription factor 2(Runx 2)(P<0.01),whereas promoted the expression of osteoprotegerin(OPG)(P<0.05),and conversely,the knockdown of DANCR-201 markedly increased Runx 2 expression and reduced OPG expression(P<0.01).DANCR-201 did not affect the lipid phagocytosis and reverse cholesterol transport ability of macrophages.Conclusions:DANCR-201 may promot

关 键 词：长链非编码RNA 分化拮抗非编码RNA-201 巨噬细胞炎症 动脉粥样硬化 

分 类 号：R541.4[医药卫生—心血管疾病]