中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (7): 492-495.

• 论著 • 上一篇    下一篇

沙利度胺对HaCaT细胞分泌血管内皮细胞生长因子及肿瘤坏死因子α的影响

苏飞1,晋红中2,李峰3,舒丹4   

  1. 1. 武汉市第一医院
    2. 中国医学科学院北京协和医学院北京协和医院
    3. 北京协和医院皮肤科
    4. 北京协和医院
  • 收稿日期:2012-09-02 修回日期:2013-03-01 出版日期:2013-07-15 发布日期:2013-07-01
  • 通讯作者: 晋红中 E-mail:jinhongzhong@263.net

Effects of thalidomide on the expression and secretion of vascular endothelial growth factor and tumor necrosis factor-alpha by a human keratinocyte cell line HaCaT

  • Received:2012-09-02 Revised:2013-03-01 Online:2013-07-15 Published:2013-07-01

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摘要： 目的 探讨沙利度胺对HaCaT细胞增殖及血管内皮细胞生长因子（VEGF）和肿瘤坏死因子α（TNF-α）表达的影响。方法 体外培养的HaCaT细胞分为阴性对照组及不同浓度沙利度胺实验组；采用水溶性四氮唑法（WST-1）检测沙利度胺对HaCaT细胞增殖的影响，实时定量PCR法检测HaCaT细胞VEGF及TNF-α mRNA的表达水平，ELISA法检测HaCaT细胞VEGF及TNF-α蛋白的表达水平。采用单因素方差分析进行统计分析。结果 沙利度胺浓度在0.01 ～ 100 nmol/L之间时，可抑制VEGF mRNA（0.439 ～ 0.634，P ＜ 0.01）与蛋白（0.587 ～ 0.923，P ＜ 0.05）的表达；浓度在0.1 ～ 100 nmol/L之间时，可抑制TNF-α mRNA（0.493 ～ 0.587，P ＜ 0.05）与蛋白（0.408 ～ 0.617，P ＜ 0.01）的表达。结论 沙利度胺在一定浓度范围可抑制角质形成细胞增殖及减少VEGF及TNF-α表达。 【关键词】 角蛋白细胞； 沙立度胺； 血管内皮生长因子类； 肿瘤坏死因子α

关键词: 角蛋白细胞, 沙利度胺, 血管内皮细胞生长因子类, 肿瘤坏死因子α

Abstract: SU Fei, JIN Hong-zhong, LI Feng, SHU Dan. Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China Corresponding author: JIN Hong-zhong, Email: jinhongzhong@263.net 【Abstract】 Objective To investigate the effect of thalidomide on the proliferation of as well as the expression and secretion of vascular endothelial growth factor （VEGF） and tumor necrosis factor-alpha （TNF-α） by the human keratinocyte cell line HaCaT. Methods Cultured HaCaT cells were divided into several groups to be treated with dimethyl sulfoxide （negative control group） and various concentrations （0.01nmol/L － 100 μmol/L） of thalidomide （experimental groups） respectively for 20 to 24 hours. Subsequently, water soluble tetrazolium-1（WST-1） assay was performed to estimate cellular proliferation, real time quantitative PCR to detect the mRNA expression of VEGF and TNF-α in HaCaT cells, and enzyme-linked immunosorbent assay （ELISA） to quantify the protein expressions of VEGF and TNF-α in the culture supernatants of HaCaT cells. Statistical analysis was done by one-way analysis of variance with least significant difference post hoc test. Results The survival rate of HaCaT cells was 74.3%, 82.9% and 90.8% after 24-hour treatment with thalidomide of 100, 10 and 1 μmol/L respectively, significantly lower than that in the negative control group （100%, all P < 0.01）. A significant decrease was induced in the mRNA expression （0.439- to 0.634-fold change, all P < 0.01） and supernatant level （（0.587- to 0.923-fold change, P < 0.05） of VEGF in HaCaT cells by thalidomide of 0.01 － 100 nmol/L, as well as in the mRNA expression （0.493- to 0.587-fold change, P < 0.05） and supernatant level （0.408- to 0.617-fold change, P < 0.01） of TNF-α by thalidomide of 0.1 － 100 nmol/L. Conclusion Within a certain range of concentration, thalidomide could suppress the proliferation of, as well as the expression and secretion of VEGF and TNF-α by, HaCaT cells. 【Key words】 Keratinocytes; Thalidomide; Vascular endothelial growth factors; Tumor necrosis factor-alpha

Key words: Tumor Necrosis Factor-alpha