人胆固醇酯水解酶对泡沫细胞的影响及相关机制的研究  

Effect of Human Cholesteryl Ester Hydrolase on Foam Cells and the Corresponding Mechanisms

作　　者：关宁[1] 王超[1] 霍晓川[1] 顾立学[1] 罗俊生[1] 

机构地区：[1]辽宁医学院附属第一医院神经外科,辽宁锦州121001

出　　处：《中国医科大学学报》2014年第9期773-776,共4页Journal of China Medical University

基　　金：国家自然科学基金(81171111);辽宁省科学事业公益研究基金(2013001009)

摘　　要：目的探讨人胆固醇酯水解酶（hCEH）表达对泡沫细胞的影响及相关机制。方法利用含hCEH的慢病毒转染RAW264．7小鼠单核巨噬细胞，转染成功后巨噬细胞泡沫化。油红O染色和高效液相色谱分析hCEH对细胞内脂滴堆积情况及游离胆固醇含量变化的影响；Western blot检测ATP结合盒转运蛋白A1（ABCA1）、ATP结合盒转运蛋白G1（ABCG1）的表达情况。结果转染72h后，荧光细胞数量最多，接近50％，hCEH在细胞内大量表达；随着时间的延长，油红O染色阳性细胞数逐渐减少，细胞内总胆固醇、游离胆固醇和胆固醇酯含量逐渐下降；Western blot显示在0～8h，ABCA1和ABCG1表达逐渐增加，8h达高峰，以后逐渐减少。结论hCEH表达可以促进巨噬细胞表面ABCA1、ABCG1的表达，并且ABCA1、ABCG1之间存在一定的协同性，从而有效增加了游离胆固醇外流、减少胆固醇酯在细胞内聚积，抑制泡沫细胞形成。Objective To study the effect of human cholesteryl ester hydrolase （hCEH） on foam cells and the corresponding mechanisms. Meth- ods RAW 264.7 macmphages were transfected transiently with Lenti-hCEH-IRES-EGFP. After successfully transfection, maerophages were trans- formed to foam ceils. Oil red O staining and high performance liquid chromatography were used to evaluate the effect of hCEH on intracellular choles- terol and cholesterol esters. Western blot was employed to detect the expression of ATP-binding cassetle transporter A 1 （ABCA 1 ） and ATP-hinding cassetle transporter G1 （ ABCG1 ）. Results The highest number of fluorescent cells was observed after 72 h, the transfection rate was about 50% and the protein expression level of hCEH significantly increased. Oil red O-positive cells together with the cellular contents of total cholesterol, free cholesterol and cholesterol ester were greatly reduced along with the prolonged time. From 0 to 8 hour, Western blot showed an increased expression of ABCA 1 and ABCG1 to the summit of 8 hour, and then decreased. When one extracellular transporter such as ApoA-I mediated cholesterol efflux, ABCA 1 was significantly expressed but ABCG1 decreased alternatively. Conclusion The expression of hCEH in RAW 264.7 macrophage stimulated the expression of ABCA1 and ABCG1. There was a synergistic relationship between ABCA1 and ABCG1 protein expression, and therefore inhibits atherosclerotic foam cell formation through increasing cholesterol efllux and reducing intracellular cholesterol level.

关 键 词：人胆固醇酯水解酶 ATP结合盒转运蛋白A1 ATP结合盒转运蛋白G1 游离胆固醇 泡沫细胞 

分 类 号：R651.1[医药卫生—外科学]