| PCSK9 siRNA对THP-1源性巨噬细胞CD36?SR-A1及SR-B1表达的影响 |
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| 引用本文： | 唐志晗,武春艳,谢 闵,刘录山,姜志胜.PCSK9 siRNA对THP-1源性巨噬细胞CD36?SR-A1及SR-B1表达的影响[J].南京医科大学学报,2011(5):673-678. |
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| 作者姓名： | 唐志晗 武春艳 谢 闵 刘录山 姜志胜 |
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| 作者单位： | 南华大学心血管病研究所,动脉硬化学湖南省重点实验室,湖南 衡阳 421001;南华大学心血管病研究所,动脉硬化学湖南省重点实验室,湖南 衡阳 421001;南华大学心血管病研究所,动脉硬化学湖南省重点实验室,湖南 衡阳 421001;南华大学心血管病研究所,动脉硬化学湖南省重点实验室,湖南 衡阳 421001;南华大学心血管病研究所,动脉硬化学湖南省重点实验室,湖南 衡阳 421001 |
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| 基金项目： | 湖南省应用基础研究计划重点项目（2008FJ2006)；湖南省科技厅计划项目（2009TP4057-2,2010TP4008-2);湖南省教育厅重点科研项目（10A105);湖南省高校科技创新团队支持计划资助 |
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| 摘 要： | 目的:研究前蛋白转换酶枯草溶菌素9(PCSK9) siRNA对THP-1源性巨噬细胞CD36?SR-A1及SR-B1表达的影响?方法:以THP-1源性巨噬细胞为研究对象,应用Lipofectamine 2000转染不同浓度PCSK9 siRNA进THP-1源性巨噬细胞中,RT-PCR及Western blot筛选出最有效的siRNA,再转染入THP-1源性巨噬细胞,24 h后加入ox-LDL处理24 h,采用油红O染色检测细胞内脂质蓄积情况,RT-PCR分析细胞CD36?SR-A1及SR-B1表达?结果:浓度为80 nmol/L的PCSK9 siRNA基因沉默效应最佳;油红O染色结果表明ox-LDL组细胞内脂质蓄积情况最为明显,PCSK9 siRNA转染组次之;PCSK9 siRNA转染组CD36 mRNA表达水平相对于ox-LDL组降低(P < 0.05),而ox-LDL组和PCSK9 siRNA转染组SR-A1及SR-B1 mRNA表达无明显差异?结论:PCSK9可能通过影响摄取脂质的细胞膜表面受体CD36表达,参与动脉粥样硬化的发生发展?
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| 关 键 词： | 前蛋白转换酶枯草溶菌素9 巨噬细胞 CD36 SR-A1 SR-B1 |
| 收稿时间： | 2010/11/19 0:00:00 |
Effects of PCSK9 siRNA on CD36,SR-A1 and SR-B1 expression in THP-1 derived macrophages |
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| TANG Zhi-han,WU Chun-yan,XIE Min,LIU Lu-shan and JIANG Zhi-sheng.Effects of PCSK9 siRNA on CD36,SR-A1 and SR-B1 expression in THP-1 derived macrophages[J].Acta Universitatis Medicinalis Nanjing,2011(5):673-678. |
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| Authors: | TANG Zhi-han WU Chun-yan XIE Min LIU Lu-shan and JIANG Zhi-sheng |
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| Institution: | Institute of Cardiovascular Disease,Key Lab for Arteriosclerology of Hunan Povince,University of South China,Hengyang 421001,China;Institute of Cardiovascular Disease,Key Lab for Arteriosclerology of Hunan Povince,University of South China,Hengyang 421001,China;Institute of Cardiovascular Disease,Key Lab for Arteriosclerology of Hunan Povince,University of South China,Hengyang 421001,China;Institute of Cardiovascular Disease,Key Lab for Arteriosclerology of Hunan Povince,University of South China,Hengyang 421001,China;Institute of Cardiovascular Disease,Key Lab for Arteriosclerology of Hunan Povince,University of South China,Hengyang 421001,China |
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| Abstract: | Objective:To investigate the effects of PCSK9 siRNA on CD36,SR-A1 and SR-B1 expressions in THP-1 derived macrophages. Methods:The siRNA for PCSK9 gene were transfected into THP-1 derived macrophages using positive ion liposome Lipofectamine 2000. Transfection efficiency was assessed by fluorescence microscope assay. The expression of PCSK9 in THP-1 derived macrophages at 24 h after transfection was detected by RT-PCR and Western blot. The most efficient siRNA was selected for transfection. At 24 h after transfection,macrophages were treated with ox-LDL for another 24 h. Then the intracellular lipid accumulation was observed by oil red O staining,and expressions of CD36 mRNA,SR-A1 mRNA and SR-B1 mRNA were detected by RT-PCR. Results:siRNA of 80 nmol/L showed the strongest inhibition for the gene expression of PCSK9,and ox-LDL-induced accumulation of cholesterol and upregulation of CD36 mRNA expression in THP-1 derived macrophages decreased by application of PCSK9 siRNA(P < 0.05). However,PCSK9 siRNA caused no effect on ox-LDL-induced upregulation of SR-A1 mRNA and SR-B1 mRNA expression in macrophages. Conclusion:PCSK9 may be involved in atherosclerosis development by inhibiting the expression of CD36. |
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| Keywords: | PCSK9 macrophages expression CD36 SR-A1 SR-B1 |
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