中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (15): 2805-2810.doi: 10.3969/j.issn.1673-8225.2012.15.032

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

小干扰RNA下调cyclophilin-D蛋白表达对内皮细胞抗氧化损伤的作用☆

彭江洲1，杨艳旗2，陈  炬2，陈柏深1，薛  雷1   

  1. 1南方医科大学第三附属医院胸心外科，广东省广州市  510630；2中山大学附属第二医院胸心外科，广东省广州市    510120
  • 收稿日期:2011-11-03 修回日期:2012-01-17 出版日期:2012-04-08 发布日期:2012-04-08
  • 通讯作者: 杨艳旗，教授，博士生导师，中山大学附属第二医院胸心外科，广东省广州市 510120 yangyanqisw@163.com
  • 作者简介:彭江洲☆，男，1975年生，湖南省吉首市人，土家族，2009年中山大学毕业，博士，主治医师，主要从事冠心病的综合治疗。 jiangzhou83@163.com

Effect of small interfering RNA down-regulated Cyclophilin-D protein expression against oxidative injury to endothelial cells

Peng Jiang-zhou1, Yang Yan-qi2, Chen Ju2, Chen Bo-shen1, Xue Lei1   

  1. 1Department of Cardiothoracic Surgery, the Third Affiliated Hospital of Southern Medical University, Guangzhou  510630, Guangdong Province, China; 2Department of Cardiothoracic Surgery, the Second Affiliated Hospital of Sun Yat-sen University, Guangzhou  510120, Guangdong Province, China
  • Received:2011-11-03 Revised:2012-01-17 Online:2012-04-08 Published:2012-04-08
  • Contact: author: Yang Yan-qi, Professor, Doctoral supervisor, Department of Cardiothoracic Surgery, the Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China yangyanqisw@163.com
  • About author:Peng Jiang-zhou☆, Doctor, Attending physician, Department of Cardiothoracic Surgery, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, Guangdong Province, China jiangzhou83@163.com

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摘要：

背景：线粒体基质中的Cyclophilin-D蛋白(CypD)在线粒体损伤过程中起着特殊作用。目前对于CypD蛋白与血管内皮细胞功能的关系，以及CypD蛋白在动脉粥样硬化和血管狭窄发生、发展中的作用仍未明确。
目的：观察小干扰RNA基因沉默对内皮细胞CyPD蛋白表达及氧化应激损伤、凋亡的影响。
方法：采用含500 μmol/L H2O2的PBS液处理ECV304细胞，小干扰RNA沉默ppif基因以建立CyPD蛋白缺陷型细胞凋亡模型，空白对照组以单纯PBS液处理。流式细胞仪检测ECV304细胞凋亡率，电镜观察细胞超微结构。
结果与结论：CyPD缺陷模型组细胞凋亡率为(32.51±6.6)%，明显低于500 μmol/L H2O2处理组(52.57±5.84)%，P=0.001。电镜观察结果显示，CyPD缺陷型细胞模型组细胞凋亡数量明显较500 μmol/L H2O2处理组减少，形态改变也较轻，多为早期凋亡特征。提示下调CyPD蛋白水平可明显减轻血管内皮细胞氧化应激损伤和凋亡。
关键词：内皮细胞；氧化应激；RNA干扰；cyclophilin-D蛋白；线粒体缩
缩略语注释：siRNA: small interfering RNA，小干扰RNA
doi:10.3969/j.issn.1673-8225.2012.15.032

关键词: 内皮细胞, 氧化应激, RNA干扰, cyclophilin-D蛋白, 线粒体缩

Abstract:

BACKGROUND: Cyclophilin-D protein in the mitochondrial matrix plays a special role in the process of mitochondrial damage. Correlation between Cyclophilin-D protein and vascular endothelial cell function, as well as Cyclophilin-D protein effects on atherosclerosis and vascular stenosis are still unclear.
OBJECTIVE: To investigate the relationship between Cyclophilin-D expression and oxidative stress damage to endothelial cells by small interfering RNA (siRNA)-mediated gene silencing.
METHODS: ECV304 cells were cultured in phosphate buffered saline containing 500 μmol/L H2O2. siRNA silenced ppif gene was used to establish Cyclophilin-D deficiency models. The blank group was treated with phosphate buffered saline. Apoptotic rate of ECV304 cells was detected by flow cytometry; meanwhile, morphological changes were observed by electron microscope.
RESULTS AND CONCLUSION: The apoptotic rate of ECV304 cells was (32.51±6.6)% in the Cyclophilin-D deficiency group, significantly lower than (52.57±5.84)% in the H2O2 group (P=0.001). The electron microscope results showed that the number of apoptotic cells in the Cyclophilin-D deficiency group was decreased significantly as compared with the H2O2 group, and the morphological changes in the former group were slighter that were characterized as early apoptosis. Cyclophilin-D is a key protein to vascular endothelial cells against oxidative stress injury.

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