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  • 付文波,李大主,丁世芳,何少林,王妍,黎明,冯义柏.氧化型低密度脂蛋白对人THP1细胞 Notch1表达及分泌细胞因子的影响[J].第二军医大学学报,2012,33(2):140-144    [点击复制]
  • FU Wen-bo,LI Da-zhu,DING Shi-fang,HE Shao-lin,WANG Yan,LI Ming,FENG Yi-bai.Effect of oxidized low-density lipoprotein on Notch1 expression and cytokine secretion in THP1 cells[J].Acad J Sec Mil Med Univ,2012,33(2):140-144   [点击复制]
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氧化型低密度脂蛋白对人THP1细胞 Notch1表达及分泌细胞因子的影响
付文波1,2,李大主1,丁世芳2,何少林1,王妍1,黎明1*,冯义柏1*
0
(1.华中科技大学协和医院心内科,华中科技大学心血管病研究所,武汉 430022
2.广州军区武汉总医院心内科,武汉 430070
*通信作者)
摘要:
目的 探讨氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)刺激人单核细胞株THP1对Notch1表达及分泌细胞因子的影响,探讨其对动脉粥样硬化(atherosclerosis,AS)发病的可能作用机制。方法 将人单核细胞株(THP1)经氟波酯(PMA)刺激转化为巨噬细胞后给予不同浓度ox-LDL刺激,相差显微镜动态观察细胞形态变化,RT-PCR检测不同浓度ox-LDL刺激后细胞表面Notch1 mRNA水平,采用Western印迹测定Notch1蛋白表达,采用ELISA法测定上清液中血管细胞黏附分子-1(vascular cell adhesive molecule-1,VCAM-1)和单核细胞趋化分子-1(monocyte chemoattractant protein-1,MCP-1)浓度。结果 ox-LDL诱导48 h后巨噬细胞形态发生树突样细胞改变; 与对照组相比,ox-LDL能刺激巨噬细胞表面Notch1蛋白和mRNA表达升高(P<0.05),培养上清液中VCAM-1和MCP-1表达升高(P<0.05),在50 mg/L浓度下诱导最佳。结论 ox-LDL能够刺激THP1细胞诱导Notch1表达增加,同时能增加动脉粥样硬化相关细胞因子VCAM-1和MCP-1的分泌,ox-LDL致动脉粥样硬化作用可能部分由Notch1介导。
关键词:  动脉粥样硬化  氧化型LDL脂蛋白类  Notch1  血管细胞黏附分子1  单核细胞趋化蛋白1
DOI:10.3724/SP.J.1008.2012.0140
投稿时间:2011-10-17修订日期:2011-12-22
基金项目:国家自然科学基金(81172781).
Effect of oxidized low-density lipoprotein on Notch1 expression and cytokine secretion in THP1 cells
FU Wen-bo1,2,LI Da-zhu1,DING Shi-fang2,HE Shao-lin1,WANG Yan1,LI Ming1*,FENG Yi-bai1*
(1. Institute of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei, China
2. Department of Cardiology, Wuhan General Hospital, PLA Guangzhou Military Area Command, Wuhan 430070, Hubei, China
*Corresponding authors.)
Abstract:
Objective To explore the effect of oxidized low-density lipoprotein (ox-LDL) on Notch 1 expression and cytokine secretion in human acute monocytic leukemia cell line THP1, so as to discuss the role of ox-LDL in atherosclerosis (AS). Methods THP1 cells were stimulated with PMA to induce human macrophages, which were then exposed to ox-LDL of different concentrations. The morphological changes of the cells was observed by phase contrast microscope. The expression of Notch1 mRNA was examined by real-time quantitative PCR (RT-PCR) and protein expression was measured by Western blotting analysis in the cells.The levels of vascular cell adhesive molecule-1(VCAM-1) and monocyte chemoattractant protein-1(MCP-1) in supernatants were determined by ELISA. Results The macrophages differentiated into dendritic like cells 48 h after ox-LDL treatment. Ox-LDL treatment increased Notch1 mRNA and protein expression compared with the control group. Ox-LDL also significantly increased the levels of VCAM-1 and MCP-1 in the supernatant in a dose-dependent manner(P<0.05). The best effect of ox-LDL was seen at the concentration of 50 mg/L. Conclusion Ox-LDL stimulation can increase Notch 1 expression in THP1 cells; meanwhile, it can also promote the secretion of AS-related cytokines. The role of ox-LDL in AS might be partly mediated by Notch1.
Key words:  atherosclerosis  oxidized LDL lipoproteins  Notch1  vascular cell adhesion molecule-1  monocyte chemotactic protein-1