中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (37): 6947-6950.doi: 10.3969/j.issn.2095-4344.2012.37.022

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

肿瘤坏死因子a促进人冠状动脉内皮细胞的凋亡

何战斌1，陈广伟2   

  1. 1开封市第一人民医院心内二科，河南省开封市 475004
    2河南大学医学院，河南省开封市 475004
  • 收稿日期:2012-01-06 修回日期:2012-02-29 出版日期:2012-09-09 发布日期:2012-09-09
  • 通讯作者: 陈广伟，硕士，河南大学医学院，河南省开封市 475004 guangweichen2009@163.com
  • 作者简介:何战斌，男，1973年生，河南省开封市人，主治医师，主要从事心血管内科临床工作。

Tumor necrosis factor alpha promotes the apoptosis of human coronary artery endothelial cells

He Zhan-bin1, Chen Guang-wei2   

  1. 1Second Department of Cardiology, the First People's Hospital of Kaifeng, Kaifeng 475004, Henan Province, China
    2 Medical College of Henan University, Kaifeng 475004, Henan Province, China
  • Received:2012-01-06 Revised:2012-02-29 Online:2012-09-09 Published:2012-09-09
  • Contact: Chen Guang-wei, Master, Medical College of Henan University, Kaifeng 475004, Henan Province, China guangweichen2009@163.com
  • About author:He Zhan-bin, Attending physician, Second Department of Cardiology, the First People's Hospital of Kaifeng, Kaifeng 475004, Henan Province, China

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440

被引次数

2

摘要：

背景：研究表明冠状动脉内皮细胞凋亡参与了动脉粥样硬化的发生、发展过程。
目的：观察肿瘤坏死因子a对人冠状动脉内皮细胞的诱导损伤作用。
方法：取对数生长期的人冠状动脉内皮细胞c-12221，分别加入含0(对照)，200，400，600 mg/L肿瘤坏死因子a的培养液，采用MTT检测细胞增殖率变化，Hoechst 33258/PI双染观察凋亡细胞形态变化，Annexin V-FITC和PI双染流式检测细胞凋亡率变化，高内涵活细胞成像系统检测细胞线粒体膜电位变化。
结果与结论：肿瘤坏死因子a呈剂量依赖性抑制人冠状动脉内皮细胞的增殖。Hoechst 33258/PI染色观察可见凋亡细胞染色质凝集、细胞核碎裂成碎片等典型细胞凋亡的特征性变化，不同质量浓度肿瘤坏死因子a组细胞凋亡率高于对照组(P < 0.05)，线粒体膜电位低于对照组(P < 0.05)，且呈剂量依赖性。表明肿瘤坏死因子a呈剂量依赖性促进人冠状动脉内皮细胞凋亡，抑制其增殖，作用机制与线粒体凋亡通路有关。

关键词: 人冠状动脉内皮细胞, 凋亡, 线粒体膜电位, 肿瘤坏死因子a, 增殖

Abstract:

BACKGROUND: Studies have shown that the apoptosis of human coronary artery endothelial cells is involved in the generation and development of atherosclerosis.
OBJECTIVE: To investigate the inductive effect of tumor necrosis factor alpha on the damage of human coronary artery endothelial cells.
METHODS: First, human coronary artery endothelial cells c-12221 in the logarithmic growth phase were selected. Next, the cells were added into 0 (control), 200, 400 and 600 mg/L tumor necrosis factor alpha culture medium. Then, the changes of cell proliferation rate, morphological changes of apoptosis cells, changes of cell apoptosis rate and mitochondrial membrane potential were detected by MTT assay, Hoechst 33258/propidium iodide staining, fluorescence microscopy, Annexin V-FITC and PI-double staining flow cytometry assay and high content live cell imaging system, respectively.
RESULTS AND CONCLUSION: MTT results showed that tumor necrosis factor alpha had potential inhibitory effects on human coronary artery endothelial cells proliferation in a dose-dependent manner. Hoechst 33258/propidium iodide staining indicated that characteristic change of apoptotic cells could be seen clearly, such as chromatin condensation, nuclear fragmentation into pieces. Besides, cell apoptosis rate of tumor necrosis factor alpha with different concentrations was higher than that of 0 mg/L tumor necrosis factor alpha (P < 0.05); their mitochondrial membrane potential was lower than that of the control and in a dose-dependent manner (P < 0.05). These results suggest that tumor necrosis factor alpha can promote the apoptosis of human coronary artery endothelial cells in a dose- and time-dependent manner and inhibit human coronary artery endothelial cell proliferation. The underlying mechanisms are related to mitochondrial apoptosis pathway.

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