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OxLDL诱导人血管内皮细胞及平滑肌细胞DNA加合物生成
引用本文:杨世明,王颖,陈莉莉.OxLDL诱导人血管内皮细胞及平滑肌细胞DNA加合物生成[J].中国组织化学与细胞化学杂志,2010,19(4):385-389.
作者姓名:杨世明  王颖  陈莉莉
作者单位:1. 华中科技大学同济医学院解剖学系组织胚胎学教研室
2. 华中科技大学同济医学院附属同济医院消化内科,湖北武汉,430030
3. 华中科技大学同济医学院附属同济医院综合内科,湖北武汉,430030
摘    要:目的探讨oxLDL参与动脉粥样硬化发生的可能机制。方法培养人血管内皮细胞及平滑肌细胞,以50μg/L oxLDL刺激24、48h后,收获细胞用于后续实验:①免疫组化染色检测DNA加合物εdA水平;②免疫组化方法检测细胞内4-HNE修饰蛋白;③western blot法检测细胞内4-HNE修饰蛋白水平。结果oxLDL刺激EC及SMC中DNA加合物εdA水平及4-HNE修饰蛋白水平均较未刺激细胞组明显升高。结果 oxLDL诱导的氧化应激、脂质过氧化反应及其继发的DNA损伤可能为oxLDL参与动脉粥样硬化发生的重要机制。

关 键 词:动脉粥样硬化  氧化低密度脂蛋白  脂质过氧化作用  亚乙烯基-DNA加合物  人内皮细胞和平滑肌细胞

OXIDIZED LOW DENSITY LIPOPROTEIN INDUCES THE FORMATION OF DNA ADDUCTS IN HUMAN VASCULAR ENDoTHELIAL AND SMOOTH MUSCLE CELLS
Yang Shiming,Wang Ying,Chen Lili.OXIDIZED LOW DENSITY LIPOPROTEIN INDUCES THE FORMATION OF DNA ADDUCTS IN HUMAN VASCULAR ENDoTHELIAL AND SMOOTH MUSCLE CELLS[J].Chinese Journal of Histochemistry and Cytochemistry,2010,19(4):385-389.
Authors:Yang Shiming  Wang Ying  Chen Lili
Institution:(Division of Histology and Embryology,Department of Anatomy,Tongji Medical College,Huazhong University of Science and Technology;1Department of Gastroenterology;2Department of Geriatrics,Affiliated Tongji Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
Abstract:Objective To investigate the pathogenesis of oxidized LDL(oxLDL) in the progression of atherosclerosis.Methods To correlate oxLDL and macromolecular damage,we measured levels of LPO-derived miscoding etheno-DNA adducts and LPO-modified proteins in cultured human vascular endothelial and smooth muscle cells after incubation with oxLDL for up to 48h.A semi-quantative analysis method for 1,N6-etheno-deoxyadenosine(εdA) by immunohistochemistry was applied.Results After oxLDL stimulation,a marked and significant increase of εdA-stained nuclei was found in both endothelial and smooth muscle cells.Similarly,4-hydroxy-2-nonenal– modified proteins,as analyzed by immunohistochemistry and Western blot,showed a 3-5 fold increase.Conclusion LPO-derived etheno-DNA adducts and LPO-modified proteins are strongly induced by oxLDL in human vascular endothelial and smooth muscle cells.This macromolecular damage may contribute to the dysfunction of arterial endothelium and the onset of atherosclerosis.
Keywords:Atherosclerosis  Oxidized LDL  Lipid peroxidation  Etheno-DNA adduct  Human endothelial and smooth muscle cells
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