同型半胱氨酸对THP-1单核细胞源性泡沫细胞形成中ABCA1、ACAT1表达的影响

检验医学 ›› 2012, Vol. 27 ›› Issue (2): 129-134.

• 临床检验与血液学检验论著 • 上一篇下一篇

同型半胱氨酸对THP-1单核细胞源性泡沫细胞形成中ABCA1、ACAT1表达的影响

1. 宁夏医科大学检验学院，宁夏银川 750004；2. 宁夏医科大学基础医学院，宁夏银川 750004

出版日期:2012-02-28 发布日期:2012-02-19
通讯作者: 姜怡邓，联系电话：0951-4086310。
作者简介:马琳娜，女，1985年生，学士，从事动脉粥样硬化与分子生物学研究。
基金资助:
国家自然科学基金资助项目（30960124）；2010年教育部新世纪优秀人才计划（NCET-10-0916）；宁夏医科大学2010大学生创新
性实验获批项目（2011[42]）

The influence of homocysteine on the expressions of ABCA1 and ACAT1 in the formation of THP-1 monocytederived foam cells

1.Medical Laboratory Science College， Ningxia Medical University, Ningxia Yinchuan 750004, China; 2. Preclinical Medicine College， Ningxia Medical University，Ningxia Yinchuan 750004，China

Online:2012-02-28 Published:2012-02-19

摘要/Abstract

摘要： 目的　探讨同型半胱氨酸(Hcy)对THP-1单核细胞源性泡沫细胞形成中三磷酸腺苷-结合转运子A1(ABCA1)和酰基辅酶A：胆固醇酰基转移酶(ACAT1)表达的影响。方法　将THP-1单核细胞与佛波酯(PMA)、氧化低密度脂蛋白(ox-LDL)共同培养，复制泡沫细胞模型，并分别用50、100、200、500 μmol/L Hcy和100 μmol/L Hcy+叶酸+维生素B₁₂(Vit B₁₂)干预72 h，并设对照组(不加入Hcy)。采用油红O染色检测泡沫细胞的形成。采用酶终点法测定细胞内总胆固醇(TC)、游离胆固醇(FC)和胆固醇酯(CE)含量的变化，观察Hcy对泡沫细胞CE流出的影响。采用荧光定量逆转录聚合酶链反应 (RT-PCR)测定ABCA1、ACAT1 mRNA表达；免疫印迹法检测ABCA1、ACAT1蛋白表达。结果　油红O染色显示Hcy加剧了泡沫细胞的形成，但不呈量效关系。实验组（加入50、100、200、500 μmol/L Hcy和100 μmol/L Hcy+叶酸+Vit B₁₂）泡沫细胞阳性百分率均高于对照组(P<0.05、P<0.01)，以100 μmol/L Hcy组效应最为明显(P<0.01)。在Hcy的干预下，泡沫细胞胞内TC、FC、CE流出减少，与对照组比较差异有统计学意义(P<0.05)，以100 μmol/L Hcy组效应最为明显(P<0.01)。100 μmol/L Hcy+叶酸+VitB₁₂组与100 μmol/L Hcy组比较，前者泡沫细胞形成减少，胆固醇流出增多。RT-PCR结果显示ABCA1 mRNA表达下调，ACAT1 mRNA表达上调，均以100 μmol/L Hcy组效应最为明显(P<0.01)；免疫印迹法检测ABCA1、ACAT1蛋白的表达与其mRNA表达一致。结论　Hcy下调了ABCA1的表达，上调了ACAT1的表达，促使了泡沫细胞形成。

关键词: 同型半胱氨酸, 三磷酸腺苷-结合转运子A1, 酰基辅酶A：胆固醇酰基转移酶

Abstract: Objective　To investigate the influence of homocysteine(Hcy) on the expressions of ATP-binding cassette transporter A1(ABCA1) and acylcoenzyme A: cholesterol acyltransferase-1 (ACAT1) in the formation of THP-1 monocytederived foam cells. Methods　THP-1 monocytes were cultured with phorbol myristate acetate(PMA) and oxidized low density lipoprotein(ox-LDL) so as to copy foam cell model， and then intervened by 50，100，200 and 500 μmol/L Hcy and 100 μmol/L Hcy+folic acid+vitamin B₁₂(Vit B₁₂) for 72 h. The control group(not intervened by Hcy) was set. The formation of foam cells was analyzed by the oil red O staining. The intracellular total cholesterol(TC)，free cholesterol(FC) and cholesterol ester(CE) of foam cells were measured by enzymatic end point determination in order to observe the effects of Hcy on foam cell CE efflux. Fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR) detected the expressions of ABCA1 mRNA and ACAT1 mRNA. Western blotting tested the expressions of ABCA1 protein and ACAT1 protein. Results　After the intervention of Hcy， oil red O staining showed that the formation of foam cells was increased significantly， but not in doseeffect relationship. The positive percentage of experimental group (50, 100, 200 and 500 μmol/L Hcy and 100 μmol/L Hcy+folic acid+Vit B₁₂) was higher than that of control group (P<0.05,P<0.01). The 100 μmol/L Hcy group was the most significant one(P<0.01). The outflow of foam intracellular TC，FC and CE was reduced significantly compared with control group(P<0.05), and the 100 μmol/L Hcy group was the most significant one (P<0.01). The 100 μmol/L Hcy+folic acid+Vit B₁₂ group showed that the formation of foam cells was decreased， whereas the cholesterol efflux was increased compared with 100 μmol/L Hcy group. RT-PCR showed that the expression of ABCA1 mRNA was decreased, but ACAT1 mRNA was increased，and the 100 μmol/L Hcy group was the most significant effect one(P<0.01). Western blotting showed that the expressions of ABCA1 protein and ACAT1 protein were consistent with mRNA expression. Conclusions　Hcy reduces the expression of ABCA1，in the contrast，the expression of ACAT1 is increased. Both of them promote the formation of foam cells.

Key words: Homocysteine, ATPbinding cassette transporter A1, Acylcoenzyme A: cholesterol acyltransferase-1

马琳娜;梁宇;于海娇;徐支芳;王艳华;姜怡邓. 同型半胱氨酸对THP-1单核细胞源性泡沫细胞形成中ABCA1、ACAT1表达的影响[J]. 检验医学, 2012, 27(2): 129-134.

MA Linna;LIANG Yu;YU Haijiao;XU Zhifang;WANG Yanhua;JIANG Yideng. The influence of homocysteine on the expressions of ABCA1 and ACAT1 in the formation of THP-1 monocytederived foam cells[J]. , 2012, 27(2): 129-134.

参考文献

［1］Chung S， Gebre AK， Seo J， et al. A novel role for ABCA1 generated large prebeta migrating nascent HDL in the regulation of hepatic VLDL triglyceride secretion［J］. J Lipid Res， 2009， 51 (4) : 729 742.

［2］　姚晓敏，宋保亮，王灿华，等. 人酰基辅酶A：胆固醇酰基转移酶（ACAT）［J］.上海交通大学学报（农业科学版），2006， 24(1):108115.

［3］　刘守钾，杨智承，刘　苑，等. 动脉粥样硬化中单核细胞招募与泡沫细胞形成［J］. 中国动脉硬化杂志，2009，17(11):957960.

［4］　车至香，李　枫，柳欣绮，等. 同型半胱氨酸在冠心病中的临床意义［J］. 检验医学,2008，23(1):37.

［5］　Paul A, Chang BH, Li L，et al. Deficiency of adipose differentiationrelated protein impairs foam cell formation and protects against atherosclerosis［J］. Circ Res， 2008， 102(12): 14921501.

［6］　Friso S， Pizzolo F， Choi SW， et al. Epigenetic control of 11 betahydroxysteroid dehydrogenase 2 gene promoter is related to human hypertension［J］. Atherosclerosis， 2008， 199(2): 323327.

［7］　Reza JZ, Doosti M, Salehipour M，et al. Modulation peroxisome proliferators activated receptor alpha(PPAR alpha) and acyl coenzyme A: cholesterol acyltransferase1(ACAT1)gene expression by fatty acids in foam cell［J］. Lipids Health Dis， 2009， 8:38.

［8］　Kumar R, McClain D, Young R， et al. Cholesterol transporter ATPbinding cassette A1 (ABCA1) is elevated in prion disease and affects PrPC and PrPSc concentrations in cultured cells［J］. J Gen Virol， 2008， 89(Pt 6): 15251532.

［9］　李建华，郭志刚，吴平生，等. 血管内皮细胞中ABCA1的表达及其在动脉粥样硬化发生中的意义［J］. 第一军医大学学报， 2004， 24(9): 980983.

［10］　Cizek SM, Bedri S, Talusan P， et al. Risk factors for atherosclerosis and the development of preatherosclerotic intimal hyperplasia［J］. Cardiovasc Pathol， 2007， 16(6): 344350.

［11］　Brunham LR, Kruit JK, Iqbal J， et al. Intestinal ABCA1 directly contributes to HDL biogenesis in vivo［J］. J Clin Invest， 2006， 116(4): 10521062.

［12］　Pasdar A, Yadegarfar G, Cumming A， et al. The effect of ABCA1 gene polymorphisms on ischaemic stroke risk and relationship with lipid profile［J］. BMC Med Genet， 2007，8:30.

［13］　YvanCharvet L, Welch C, Pagler TA， et al. Increased inflammatory gene expression in ABC transporterdeficient macrophages: free cholesterol accumulation， increased signaling via tolllike receptors, and neutrophil infiltration of atherosclerotic lesions［J］. Circulation， 2008, 118(18): 18371847.

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同型半胱氨酸对THP-1单核细胞源性泡沫细胞形成中ABCA1、ACAT1表达的影响

The influence of homocysteine on the expressions of ABCA1 and ACAT1 in the formation of THP-1 monocytederived foam cells

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