白细胞介素22对HaCaT细胞CCL28表达的影响  

Effects of Interleukin-22 on the Expression of CCL28 in HaCaT Cell

作　　者：熊传茜 周文娇[1] 王惠平[1] 韩珑[1] 樊雁飞 刘原君[1] 刘全忠[1] 罗素菊[1] XIONG Chuanxi;ZHOU Wenjiao;WANG Huiping;HAN Long;FAN Yanfei;LIU Yuanjun;LIU Quanzhong;LUO Suju(Department of Dermatology,Tianjin Medical University General Hospital,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院皮肤性病科,天津300052

出　　处：《中国皮肤性病学杂志》2020年第4期383-387,共5页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金(81773319)。

摘　　要：目的探讨白细胞介素(IL-22)对HaCaT细胞黏膜相关上皮趋化因子(CCL28)表达的影响。方法培养HaCaT细胞,将其分为6组:4个IL-22组(分别用12.5、25、50、100μg/L的IL-22进行干预处理);阻断剂组(50μmol/L PD98059阻断干预,2 h后加入50μg/L IL-22);对照组(用PBS处理)。24 h后,用CCK-8检测细胞的增殖;用实时荧光定量RT-PCR检测CCL28 mRNA的水平变化;用蛋白免疫印迹法、酶联免疫吸附法和免疫荧光检测CCL28蛋白水平的变化。结果CCK-8检测显示,上述浓度的IL-22作用于HaCaT细胞24 h后,对细胞的增殖有明显的促进作用,且这种促进作用能被PD98059抑制,差异具有统计学意义(P<0.05)。实时荧光定量RT-PCR检测显示,上述浓度的IL-22作用于HaCaT细胞,细胞中CCL28 mRNA逐渐升高,均较对照组升高,差异有统计学意义(P<0.01);通路阻断剂组较50μg/L IL-22组降低,差异有统计学意义(P<0.05)。用蛋白免疫印迹法和酶联免疫法检测到CCL28蛋白水平变化的趋势与实时荧光定量RT-PCR检测到的CCL28 mRNA的水平变化的趋势一致,且差异有统计学意义(P<0.05)。免疫荧光检测显示,HaCaT细胞内CCL28蛋白主要表达在细胞质。结论IL-22可剂量依赖促进HaCaT细胞增殖和CCL28的表达,其可能通过MAPK-ERK1/2通路作用。Objective To evaluate the effects of interleukin-22(IL-22) on the expression of CCL28 in HaCaT cells.Methods Cultured HaCaT cells were divided into 6 groups:IL-22 treated groups(treated with 12.5,25,50 and 100 μg/L IL-22 respectively),inhibition group(treated with 50 μmol/L PD98059 for 2 hours followed by the treatment with 50 μg/L IL-22),control group(treated with PBS).After 24-hour treatment,proliferation of HaCaT cell was determined by CCK8,the mRNA level of CCL28 were detected by real-time fluorescence-based quantitative PCR,the protein expression of CCL28 were detected by Western blot,enzyme-linked immunosorbent assay and immunofluorescence assay.Results CCK8 assay results showed that different concentrations of interleukin 22 exerted a significant promoting effect on HaCaT cell proliferation,and the promoting effect can be reversed by PD98059.The real-time quantitative RT-PCR results showed that the mRNA level of CCL28 gradually increased after IL-22 treatment in different concentrations,which were significantly higher than that in the control group(P<0.01).The level of CCL28 mRNA treated with PD98059 was lower than 50 μg/L IL-22 group,and the difference was statistically significant(P<0.05).Western blot and enzyme-linked immunosorbent assay results showed that the protein level changes of CCL28 were consistent with the changes of CCL28 mRNA.The immunofluorescence assay showed that CCL28 protein was mainly expressed in the cytoplasm of HaCaT cells.Conclusion IL-22 can promote the expression of CCL28 in HaCaT cells on a dose-dependently manner,which may be regulated the MAPK-ERK1/2 signaling pathways.

关 键 词：细胞介素22 银屑病 丝裂原激活蛋白激酶类 CCL28 HACAT细胞 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]