中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (7): 501-504.

• 论著 • 上一篇    下一篇

肿瘤坏死因子-α、白介素-6和干扰素γ对HaCaT细胞表达CD68抗原的影响

郭英军1,肖汀2,王雅坤3,陈洪铎4,赵玉铭3   

  1. 1. 沈阳市中国医科大学附属盛京医院皮肤科
    2. 中国医科大学附属第一医院皮肤科
    3. 沈阳 中国医科大学附属第一医院皮肤科
    4. 中国医科大学第一附属医院皮肤科
  • 收稿日期:2010-08-24 修回日期:2011-01-23 出版日期:2011-07-15 发布日期:2011-07-12
  • 通讯作者: 赵玉铭 E-mail:ymzhaodc05@yahoo.com
  • 基金资助:

    正常人真皮中免疫细胞分布规律和相互作用机制的研究

Effects of tumor necrosis factor （TNF）-α, interleukin （IL）-6 and interferon （IFN）-γ on the expression of CD68 in HaCaT cells

XIAO Ting2, 2,CHEN Hong-Duo 2,   

  • Received:2010-08-24 Revised:2011-01-23 Online:2011-07-15 Published:2011-07-12

PDF

35

赞

摘要：

目的 研究肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）和干扰素γ（IFN-γ）以及细菌脂多糖（LPS）对CD68抗原在角质形成细胞株HaCaT细胞中表达的影响。方法 RPMI 1640培养液培养的HaCaT细胞随机分成自然增殖孔（无刺激组）、IFN-γ刺激孔（组）、TNF-α刺激孔（组）、LPS刺激孔（组）、IL-6刺激孔（组）。培养24 h后收集细胞，采用流式细胞仪、免疫组化和RT-PCR方法检测TNF-α、IL-6、IFN-γ和LPS作用HaCaT细胞后CD68表达情况。结果 与无刺激组比较，不同刺激因素可使HaCaT细胞CD68阳性细胞数增多，但以TNF-α和IL-6的作用较强（t值为3.60和3.93，P值均 < 0.01），IFN-γ和LPS的作用稍弱（t值为2.38和2.52，P值均 < 0.05）。只有IL-6可以使HaCaT细胞CD68阳性细胞平均荧光强度增强（t值为8.34，P值 < 0.01）。在IFN-γ和TNF-α、IL-6作用HaCaT细胞24 h后均有CD68表达于胞质和胞膜，并且以TNF-α和IL-6作用较强。TNF-α和IL-6作用HaCaT细胞24 h后可见CD68 mRNA表达明显增强（t值为4.34和7.52，P值均 < 0.01），IFN-γ作用后出现较弱表达（t = 2.81，P < 0.05）。结论 在IL-6、TNF-α、IFN-γ和LPS的作用下HaCaT细胞CD68表达增强。

关键词: 流式细胞仪

Abstract:

Objective To study the effects of some cytokines such as TNF-α，IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells. Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group （without stimulation）, IFN-γ-stimulated group, TNF-α-stimulated group, LPS-stimulated group and IL-6 stimulated group. The work concentration of TNF-α, IL-6, IFN-γ and LPS was 50 mg/L. HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry, immunohistochemistry and reverse transcription （RT）-PCR, respectively. Results Compared with untreated HaCaT cells, the count of CD68-positive cells was elevated in cells stimulated by TNF-α （t = 3.60, P < 0.01）, IL-6 （t = 3.93, P < 0.01）, IFN-γ （t = 2.38, P < 0.05） and LPS （t = 2.52, P < 0.05）, and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS. Among the four cytokines, only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells （t = 8.34, P < 0.01）. After 24-hour treatment with TNF-α, IFN-γ and IL-6, CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells, and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines. A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α （t = 4.34, P < 0.01）, IL-6 （t = 7.52, P < 0.01） and IFN-γ （t = 2.81, P < 0.05）; TNF-α and IL-6 showed a stronger promotive effect than IFN-γ. Conclusion IL-6, TNF-α, IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

Key words: flow cytometric analysis