NO/PKG对THP-1巨噬细胞ABCA1基因mRNA表达和胆固醇含量的影响  

The effect of NO/PKG on ABCA1 mRNA expression and cholesterol content in THP-1 momocyte-derived macrophages

作　　者：李帅[1] 黄健[1] 李红艳[1] 莫新玲[2] 左晓娜[1] 

机构地区：[1]桂林医学院生物技术学院生物化学教研室,广西桂林541004 [2]桂林医学院附属医院心血管内科

出　　处：《临床心血管病杂志》2011年第10期791-794,共4页Journal of Clinical Cardiology

基　　金：广西自然科学基金资助项目(No:2010GXNSFA013254);广西高校百名中青年学科带头人资助计划项目(No:RC20060710016)

摘　　要：目的:探讨THP-1巨噬细胞泡沫化过程中NO/PKG的变化,以及NO/PKG对THP-1巨噬细胞ATP结合盒转运体A1(ABCA1)基因mRNA表达和胆固醇含量的影响。方法:THP-1细胞诱导分化为巨噬细胞,用氧化低密度脂蛋白(ox-LDL)处理,油红"O"染色观察巨噬细胞脂滴,酶标仪测定一氧化氮(NO)的释放;用一氧化氮供体L-精氨酸和硝普钠(SNP)、乙二醇二乙醚二胺四乙酸(EGTA)、PKG激动剂处理巨噬细胞,RT-PCR法测定细胞ABCA1基因mRNA的表达,高效液相色谱法(HPLC)测定细胞内胆固醇的含量变化。结果:ox-LDL可导致THP-1巨噬细胞脂质的蓄积和NO释放的增加,实验组NO释放量较对照组显著升高(P<0.05),实验组中50mg/L组、100mg/L组、200mg/L组与25mg/L组相比显著升高(P<0.05)。用PKG激动剂处理巨噬细胞后促进ABCA1基因mRNA表达上调(P<0.05);用PKG激动剂、L-精氨酸、SNP处理后的巨噬细胞内胆固醇较对照组显著减少(P<0.05)。EGTA能显著增加细胞内胆固醇含量(P<0.05)。结论:NO/PKG能降低细胞内胆固醇,其机制可能是PKG激活后通过上调ABCA1的表达,减少细胞内胆固醇的蓄积。Objective：To investigate the changes in NO/PKG during the process of THP-1 monocyte-derived macrophages to form foam cells and the effect of NO/PKG on macrophages ABCA1 mRNA expression and cholesterol content.Methods：THP-1 derived macrophages were incubated with ox-LDL for 48 h.Cellular lipid accumulation was determined by Oil Red O staining.Cell culture supernatant＇s total nitric oxide concentration was determined by Total Nitric Oxide Assay Kit.THP-1 derived macrophages were incubated with nitric oxide donor（SNP or L-arginine）,EGTA,PKG agonist 8-Br-cGMP,respectively.ABCA1 mRNA and cellular lipid accumulation was determined by reverse trancriptase-polymerase chain reaction（RT-PCR） and high performance liquid chromatography analysis（HPLC）,respectively.Results：Ox-LDL accumulation and increase the release of total NO（P0.05）,8-Br-cGMP can up-regulated the expression of ABCA1 mRNA.8-Br-cGMP,L-arginine,SNP could reduce the content of cellular cholesterol（P0.05）.However,EGTA increased intracellular cholesterol content.Conclusion：The NO/PKG pathway may play an important role on decreasing cellular cholesterol,the possible mechanism may be related to the up-regulation of ABCA1 mRNA expression by PKG.

关 键 词：动脉粥样硬化 PKG ABCA1 巨噬细胞 

分 类 号：R543.1[医药卫生—心血管疾病]