罗格列酮对泡沫细胞中胆固醇贮存与运输相关蛋白ACAT-1、ABCA-1表达的影响  

Effect of rosiglitazone on expression of acyl-coenzyme A cholesterol acyltransferase 1 and ATP-binding cassette transporter A1 in foam cells

作　　者：蒙颖[1] 徐芳[1] 王志禄[2] 李万玲[1] 贾军正[1] 郭文芬[1] 谢宛霞[1] 胡海英[1] 胡旭堂[1] 

机构地区：[1]兰州大学第一临床医学院心内科,兰州730000 [2]兰州大学第一医院心内科,兰州730000

出　　处：《第三军医大学学报》2012年第22期2288-2291,共4页Journal of Third Military Medical University

基　　金：甘肃省自然科学研究基金(1010RJZA122);甘肃省卫生行业科研计划管理项目(GWGL2010-16)~~

摘　　要：目的观察罗格列酮对RAW264.7巨噬源性泡沫细胞形成中胆固醇含量及酰基辅酶A-胆固醇酰基转移酶-1(acyl coenzyme A cholesterol acyl transfer enzyme 1,ACAT-1)、三磷酸腺苷结合盒转运蛋白A-1(ATP combination box oftransporter 1,ABCA-1)表达的影响,探讨罗格列酮对泡沫细胞形成的影响及可能的作用机制。方法在DMEM高糖培养基中培养RAW264.7巨噬细胞,按完全随机分组方法分为:①空白对照组(常规培养基培养巨噬细胞);②氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)组(用终浓度为30 mg/L的oxLDL孵育48 h);③oxLDL+罗格列酮组(分别用终浓度5、10、20μmol/L的罗格列酮+30 mg/L oxLDL共同孵育48 h)(n=10)。采用油红O染色观察泡沫细胞,胆固醇检测试剂盒测定各组细胞内总胆固醇(total cholesterol,TC)和游离胆固醇(free cholesterol,FC)的含量,Western blot法检测各组细胞ACAT-1和ABCA1的表达水平。结果 oxLDL组中大量泡沫细胞的胞质被油红O染色,oxLDL+罗格列酮组泡沫细胞胞质染色明显浅于oxLDL组的细胞;与oxLDL组相比,oxLDL+罗格列酮组TC及FC显著降低(P<0.05),且呈浓度依赖性;Western blot检测结果表明,不同浓度(5、10、20μmol/L)oxLDL+罗格列酮组ACAT-1蛋白表达分别为(0.94±0.11)、(0.86±0.13)、(0.58±0.12),与oxLDL组(1.19±0.12)相比有显著差异(P<0.05),不同浓度oxLDL+罗格列酮组ABCA1蛋白表达分别为(0.72±0.08)、(0.91±0.15)、(1.15±0.11),与oxLDL组(0.63±0.05)相比有显著差异(P<0.05),且呈浓度依赖性。结论罗格列酮可能通过抑制ACAT-1的表达及促进ABCA-1的表达减少泡沫细胞形成,从而发挥其抗动脉粥样硬化的作用。Objective To investigate the effect of rosiglitazone on cholesterol content and the expression of acyl-coenzyme A cholesterol acyltransferase 1（ACAT1） and ATP-binding cassette transporter A1（ABCA1） in RAW264.7 macrophage-derived foam cells,and to explore the mechanism of rosiglitazone in foam cell formation.Methods Mouse macrophage RAW264.7 cells were cultured in high-glucose DMEM and were randomly divided into a control group,an oxidized low-density lipoprotein（oxLDL） group,in which cells were incubated with 30 mg/L oxLDL for 48 h,and an oxLDL＋rosiglitazone group,in which cells were incubated with 30 mg/L oxLDL plus 5,10 and 20 μmol/L rosiglitazone for 48 h,respectively（n=10）.The formation of foam cells were identified by oil red O staining.The levels of intracellular total cholesterol（TC） and free cholesterol（FC） in each group were determined by a cholesterol detection kit,and the expression of ACAT-1 and ABCA-1 protein was determined by Western blotting.Results Mouse macrophage RAW264.7 cells in the oxLDL group were stained by oil red O,but the color of the macrophages in the oxLDL＋rosiglitazone group was lighter than that in the oxLDL group.Compared with the oxLDL group,the contents of intracellular TC and FC in the oxLDL＋rosiglitazone group significantly decreased（P0.05） in a dose-dependent manner.Western blot results showed the expression levels of ACAT-1 in the oxLDL＋rosiglitazone group with different concentrations of rosiglitazone（5,10 and 20 μmol/L） were（0.94±0.11）,（0.86±0.13） and（0.58±0.12）,respectively,which were significantly different from that in the oxLDL group（1.19±0.12）（P0.05）.The expression levels of ABCA-1 in the oxLDL＋rosiglitazone group with different concentrations of rosiglitazone（5,10 and 20 μmol/L） were（0.72±0.08）,（0.91±0.15） and（1.15±0.11）,respectively,which were also significantly different from that in the oxLDL group（0.63±0.05）（P0.05）.Conclusion Rosiglitazone may inhibit foam cell formati

关 键 词：罗格列酮 泡沫细胞 胆固醇 动脉粥样硬化 

分 类 号：R543.5[医药卫生—心血管疾病] R966[医药卫生—内科学]