白介素22调控角质形成细胞表达角蛋白17的研究

IL-22 up-regulates keratin 17 expression in human keratinocytes

目的:研究白介素(IL)-22对培养的人永生化角质形成细胞(HaCaT细胞)表达角蛋白(K)17的影响。方法:对体外培养的HaCaT细胞分别给予不同浓度的IL-22(0～100μg/L)作用24 h,应用实时荧光定量聚合酶链式反应(real-time PCR)检测K17mRNA表达水平,以酶联免疫吸附测定技术(ELISA法)、蛋白质免疫印记法(Western blot)及免疫荧光染色法检测K17蛋白表达水平的变化。结果:HaCaT细胞经12.5、25.0、50.0、100.0μg/L的IL-22作用后,均出现不同程度的K17表达。与空白对照组的HaCaT细胞相比,12.5μg/L组的mRNA及蛋白表达无明显差异,而25.0、50.0、100.0μg/L组的mRNA及蛋白表达则明显上升(P<0.05),并且随着IL-22浓度增高,K17mRNA及蛋白表达量增多。免疫荧光染色图片显示,随着IL-22浓度的增高,HaCaT细胞胞质中K17蛋白荧光染色增强。结论:IL-22可以剂量依赖方式诱导HaCaT细胞表达K17。

Objective： To investigate K17 expression affected by IL-22 in cultured human keratinocytes. Methods： HaCaT cells were treated respectively with different concentration of IL-22 （0.0 μg/L, 12.5 μg/L, 25.0 μg/L, 50.0 μg/L, and 100.0 μg/L） in vitro for 24 hours. Then K17 mRNA level was detected by real-time PCR. To further confirm the finding, K17 protein expression was detected by enzyme linked immunosorbent assay （ELISA）,Western blot and immunofluorescence staining. Results： KIT expression was observed in HaCaT cells treated with IL-22. The expression of K17 mRNA and protein elevated evidently along with the increase of IL-22 concentrations （P 〈 0.05）, while no significant change was observed in the control group or in the 12.5 μg/L group. Conclusion： IL-22 can up-regulate K17 expression in HaCaT cells in a dose-dependent manner.