摘要
目的研究同型半胱氨酸(HCY)是否通过过氧化物酶增殖物激活受体α(PPARα)-肝脏X核受体α(LXRα)通路导致动脉粥样硬化(AS)。方法RAW264.7源性巨噬细胞经氧化型低密度脂蛋白(ox-LDL)诱导为泡沫细胞,采用油红“O”染色进行泡沫细胞的鉴定。用不同浓度梯度(空白对照,0.5,1.0,2.0,5.0mmol/L)HCY干预泡沫细胞24h,油红“O”染色观察细胞内脂滴数量。Westernblot和实时定量PCR检测PPARα、LXRα及三磷酸腺苷结合盒转运体A1(ABCA1)、三磷酸腺苷结合盒转运体G1(ABCG1)、卵磷脂胆固醇酰基转移酶(LCAT)的蛋白及mRNA表达,液态闪烁计数法分析细胞内胆固醇流出率,高效液相色谱分析细胞内胆固醇酯含量。结果与空白对照组比较,HCY干预组PPARα、LXRα、ABCA1、ABCG1、LCAT的表达量均降低(P<0.05),细胞内的胆固醇含量均增高(P<0.05),细胞内胆固醇流出率降低(P<0.05)。随HCY浓度增加,PPARα、LXRα、ABCA1、ABCG1、LCAT的表达量逐渐降低(P<0.05),细胞内的胆固醇含量逐渐增高(P<0.05),细胞内胆固醇流出率逐渐降低(P<0.05)。结论HCY通过抑制PPARα-LXRα途径降低ABCA1、ABCG1、LCAT的表达,降低胆固醇逆转运效率,致脂质聚集增加,进而导致动脉粥样硬化(AS)。
Objective To investigate whether homocysteine causes atherosclerosis through the PPARα-LXRαpathway.Methods RAW264.7-derived macrophages were induced to foam cells by oxidized low density lipoprotein(ox-LDL),and oil red“O”staining was used to identify foam cells.After the foam cells were treated with different concentrations of HCY(blank control,0.5,1,2,5 mmol/L)for 24 h,the oil red“O”staining was used to observe the number of intracellular lipid droplets.Western blot and real-time quantitative PCR were used to detect the protein and gene expression of PPARα,LXRαand ATP-binding cassette transporter A1(ABCA1),ATP-binding cassette transporter G1(ABCG1),lecithin cholesterol acyltransferase(LCAT).The liquid scintillation counting was used to analyze the cholesterol efflux rate,and high performance liquid chromatography was used to analyze the intracellular cholesterol ester content.Results Compared with blank control group,the expression of PPARα,LXRα,ABCA1,ABCG1 and LCAT decreased in HCY intervention group(P<0.05),the intracellular cholesterol content increased(P<0.05),and the intracellular cholesterol efflux rate decreased(P<0.05).With the increase of HCY concentration,the expression of PPARa,LXRa,ABCA1,ABCG1 and LCAT was decreased(P<0.05),the intracellular cholesterol content was increased(P<0.05),and the intracellular cholesterol outflow rate was decreased(P<0.05).Conclusion HCY may reduce the expression of ABCA1,ABCG1 and LCAT by inhibiting PPARα-LXRαpathway,decrease the efficiency of cholesterol reverse transport and increase lipid aggregation,which leads to atherosclerosis(AS).
作者
代佩
高奋
高宏伟
王远
冯高洁
白瑞
秦卫伟
宋晓苏
李虹
DAI Pei;GAO Fen;GAO Hongwei;WANG Yuan;FENG Gaojie;BAI Rui;QIN Weiwei;SONG Xiaosu;LI Hong(Department of Paediatrics,Second Clinical Medical College,Shanxi Medical University,Taiyuan 030001,China)
出处
《山西医科大学学报》
CAS
2019年第6期695-701,共7页
Journal of Shanxi Medical University
基金
山西省自然科学基金资助项目(2014011040-3)
