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同型半胱氨酸对人脐静脉内皮细胞IκB和NF-κB表达的影响

徐倩   

  1. 承德医学院08级研究生班
  • 收稿日期:2010-09-17 修回日期:2010-11-09 出版日期:2011-05-15 发布日期:2011-05-15
  • 通讯作者: 徐倩

Effects of homocysteine on the expression of IκB and NF-κB in the human umbilical veins endothelial cells

qian XU   

  • Received:2010-09-17 Revised:2010-11-09 Published:2011-05-15 Online:2011-05-15
  • Contact: qian XU

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摘要： 目的: 观察同型半胱氨酸对人脐静脉内皮细胞NF-κB、IκB表达的影响，探讨同型半胱氨酸导致动脉粥样硬化的分子机制。方法：将体外培养的人脐静脉内皮细胞分为5组，分别加入同型半胱氨酸0mmol/L，2.5 mmol/L ,5 mmol/L,10 mmol/L ,15 mmol/L ,培养24h。通过MTT比色法测定细胞活力，采用RT-PCR法检测NF-κB p65 mRNA的表达，Western Blot检测IκB-α 的蛋白表达，免疫组化检测NF-κB p65蛋白的表达。结果：加入不同浓度的Hcy 后，HUVECs的细胞活力降低， NF-κB mRNA表达明显增强，IκB-α 蛋白表达水平显著下降，HUVECs胞核内NF-κB p65蛋白表达增强。结论：Hcy可降低HUVECs细胞存活率，通过上调NF-κB p65 mRNA的表达，增加IκB- α 蛋白的降解，促进NF-κB的核转移，进而促进NF-κB的活化。

关键词: 人脐静脉内皮细胞, 同型半胱氨酸, 四甲基噻唑氮蓝, 核因子κB, 核因子κB抑制蛋白

Abstract: Objective: to investigate the effects of homocysteine on the expression of IκB and NF-κB in the human umbilical veins endothelial cells and explore the role of homocysteine in the development of atherosclerosis. Methods:The HUVECs were randomly divided into 5 groups,and each group was cultured with 0 mmol/L Hcy,2.5 mmol/L Hcy ,5 mmol/L Hcy,10 mmol/L Hcy ,15 mmol/L Hcy for 24 hours. MTT assay was used to detect cell viability.The expression of NF-κB p65 mRNA was observed by RT-PCR, and the expression of IκB-α was observed by Western blot,and the expression of NF-κB p65 protein was observed by immunohistochemistry. Results: After HUVECs were exposed to Hcy at different concentrations, the HUVECs were inhibition. The expression of NF-κB p65 mRNA and NF-κB p65 protein were increased significantly, however, the expression of IκB-α was reduced. Conclusion: Hcy can reduce cell viability. Hcy can increase the expression of NF-κB p65 mRNA obviously, and reduce the expression of IκB-α,and increase NF-κB nuclear translocation. In a word, Hcy can increase the activity of NF-κB.

Key words: Human Umbilical Veins Endothelial Cells, homocysteine, 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, Nuclear factor κB, theinhibitor ofNFκB