NgBR对巨噬细胞源性泡沫细胞胆固醇逆转运的影响  

Effect of NgBR on Reverse Cholesterol Transport of Macrophage Derived Foam Cells

作　　者：王玉凡[1] 宋晓苏[2] 白瑞[2] 梁斌[2] 刘改珍[2] 娄秀萍[2] 尔璐 侯曜曜 张永亮[1] 边云飞[2] 肖传实[3] WANG Yu-Fan SONG Xiao-Su BAI Rui LIANG Bin LIU Gai-Zhen LOU Xiu-Ping ER Lu HOU Yao-Yao ZHANG Yong-Liang BIAN Yun-Fei and XIAO Chuan-Shi(Shanxi Medical University Department of Cardiology, the Second Hospital of Shanxi Medical University ＆ Key Labora- tory of Cardiovascular Disease Diagnosis, Treatment and Clinical Pharmacology of Shanxi Province Department of Cardiology, the First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, China)

机构地区：[1]山西医科大学 [2]山西医科大学第二医院心内科、心血管疾病诊治及临床药理山西省重点实验室 [3]山西医科大学第一医院心内科,山西省太原市030001

出　　处：《中国动脉硬化杂志》2016年第9期893-898,共6页Chinese Journal of Arteriosclerosis

基　　金：国家杰出青年科学基金项目(81400338)

摘　　要：目的通过转染小干扰RNA(siRNA)沉默RAW264.7细胞源性泡沫细胞神经轴突生长抑制因子B受体(Ng BR)表达,研究Ng BR对泡沫细胞胆固醇逆转运(RCT)的影响,探索从RCT途径抗动脉粥样硬化(As)的新方法,为冠心病的临床防治提供新思路。方法利用氧化型低密度脂蛋白(ox-LDL)诱导RAW264.7细胞形成泡沫细胞,油红O染色进行鉴定。将泡沫细胞分为四组:空白对照组、siRNA阴性对照组、Ng BR-siRNA1转染组(si Ng BR-1组)、Ng BR-siRNA2转染组(si Ng BR-2组)。利用siRNA沉默泡沫细胞Ng BR基因表达,并利用real-time PCR和Western blot对其进行干扰效率鉴定。随后采用real-time PCR检测各组细胞肝X受体α(LXRα)、三磷酸腺苷结合盒转运体A1(ABCA1)及三磷酸腺苷结合盒转运体G1(ABCG1)mRNA表达,Western blot检测各组细胞相应蛋白含量,液闪计数仪检测胆固醇流出率。结果 ox-LDL成功诱导泡沫细胞形成;si Ng BR-1组和si Ng BR-2组Ng BR mRNA及其蛋白明显下调(P<0.05);si Ng BR-1组和si Ng BR-2组LXRα、ABCA1和ABCG1的mRNA及其蛋白表达显著降低(P<0.05),胆固醇流出显著减少(P<0.05)。结论 Ng BR可以增加巨噬细胞源性泡沫细胞RCT的调控基因LXRα及其下游基因ABCA1、ABCG1的表达,从而减弱或者避免As的发生和发展,为冠心病的临床防治提供理论依据。Aim By transfection of small interfering RNA（ siRNA） to silence RAW264. 7 derived foam cells＇ neurite outgrowth inhibitor-B receptor（ Ng BR） expression,to study the effect of Ng BR on reverse cholesterol transport（ RCT） of foam cells,explore new methods to prevent atherosclerosis from RCT pathway and provide new ideas for clinical prevention and treatment of coronary heart disease. Methods Using oxidized low density lipoprotein（ ox-LDL） to induce the RAW264. 7 cells to form foam cells,and using the oil red O staining to identify them. Then the foam cells were divided into 4 groups： blank control group,siRNA negative control group,Ng BR-siRNA1 transfection group（ si Ng BR-1group） and Ng BR-siRNA2 transfection group（ si Ng BR-2 group）. Whereafter siRNA was used to silence Ng BR expression in RAW264. 7 cells,and the interference efficiency was identified by real-time PCR and Western blot. Then realtime PCR was applied to detect mRNA content of liver X receptor alpha（ LXRα）,ATP-binding cassette transporter A1（ ABCA1）,ATP-binding transporter G1（ ABCG1） in cells of each group,and corresponding protein content of each group cells were detected by Western blot,and the intracellular cholesterol efflux was determined by liquid scintillation counter.Results Ox-LDL induced foam cells formation successfully. Compared with other groups,NgB R mRNA and protein were significantly decreased in siN gB R-1 and siN gB R-2 group（ P〈0. 05）,mRNA and protein expressions of ABCA1,LXRα,and ABCG1 were significantly inhibited（ P〈0. 05）,and the cholesterol efflux was significantly reduced in siN gB R-1 and siN gBR-2 group（ P〈0. 05）. Conclusion NgB R can increase the expression of LXRα and its downstream genes as ABCA1 and ABCG1 related to RCT regulation of macrophage derived foam cells,thereby weaken or avoid the occurrence and development of atherosclerosis,and provide the theoretical basis for clinical prevention and treatment of coronary heart disease.

关 键 词：神经轴突生长抑制因子B受体 胆固醇逆转运 巨噬细胞源性泡沫细胞 

分 类 号：R363[医药卫生—病理学]