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

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

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RPMI1640培养液培养的HaCaT细胞随机分成自然增殖孔（无刺激组）、IFN一1刺激孔（组）、TNF-α刺激孔（组）、LPS刺激孔（组）、IL-6刺激孔（组）。培养24h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24h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24h后可见CD68mRNA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IL6、TNF-α、IFN-γ和LPS的作用下HaCaT细胞CD68表达增强。

Objective To study the effects of some cytokines such as TNF-α,IL-6 and IFN-~/as well as lipopolysaccharide on CD68 expression in HaCaT cells. Methods Human HaCaT keratinoeytes 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.