中华皮肤科杂志 ›› 2015, Vol. 48 ›› Issue (9): 637-640.

• 研究报道 • 上一篇    下一篇

白细胞介素22对HaCaT细胞他扎罗汀诱导基因3表达的影响

罗素菊1,刘欣欣2,郑焱3,许文娟2,李燕4,刘全忠1   

  1. 1. 天津医科大学总医院皮肤性病科
    2. 天津医科大学总医院
    3. 西安交通大学第二医院皮肤科
    4. 天津医科大学总医院皮肤科
  • 收稿日期:2014-12-10 修回日期:2015-05-04 出版日期:2015-09-15 发布日期:2015-09-01
  • 通讯作者: 罗素菊 E-mail:luosuju2005@163.com
  • 基金资助:

    国家自然科学基金;国家自然科学基金

Effects of interleukin-22 on the expression of tazarotene-induced gene 3 in HaCaT cells

  • Received:2014-12-10 Revised:2015-05-04 Online:2015-09-15 Published:2015-09-01

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摘要：

目的 探讨白细胞介素22（IL-22）对HaCaT细胞他扎罗汀诱导基因3（TIG3）表达的影响。 方法 用12.5 ~ 100 μg/L IL-22、IL-22 + PD98059（MAPK-ERK1/2通路抑制剂）及IL-22 + AG490（JAK2/STAT3通路抑制剂）干预处理HaCaT细胞24 h后，分别提取HaCaT细胞总蛋白及总RNA，用免疫荧光、Western印迹法、ELISA法检测TIG3的蛋白水平，用实时荧光定量RT-PCR检测TIG3 mRNA水平的改变。 结果 免疫荧光检测显示，HaCaT细胞内TIG3蛋白主要表达在细胞质。用Western印迹法检测，用12.5、25、50、100 μg/L的IL-22干预处理后，HaCaT细胞中TIG3蛋白表达分别为0.743 ± 0.035，0.678 ± 0.040，0.582 ± 0.041和0.328 ± 0.032，均低于对照组0.839 ± 0.045（P < 0.05）。酶联免疫法检测的结果示，上述浓度的IL-22干预处理后，TIG3蛋白水平变化的趋势与Western印迹法的结果一致。定量RT-PCR检测示TIG3 mRNA分别为对照组的0.838 ± 0.036，0.686 ± 0.061，0.565 ± 0.047，0.457 ± 0.033（P < 0.05）。加入信号通路抑制剂后，TIG3蛋白和mRNA水平降低程度较无抑制剂组减少，差异有统计学意义。 结论 IL-22可剂量依赖抑制HaCaT细胞TIG3的表达，其机制可能与MAPK-ERK1/2和JAK2/STAT3通路有关。

Abstract:

Luo Suju*, Liu Xinxin, Zheng Yan, Xu Wenjuan, Li Yan, Liu Quanzhong. *Department of Dermatology, Tianjin Medical University General Hospital, Tianjin 300052, China Corresponding author: Luo Suju, Email: luosuju2005@163.com 【Abstract】 Objective To evaluate the effects of interleukin-22 （IL-22） on the expression of tazarotene-induced gene 3 （TIG3） in HaCaT cells. Methods Cultured HaCaT cells were randomly divided into several groups to be treated with different concentrations （12.5, 25, 50, 100 μg/L） of IL-22 alone, or the combination of 50 μg/L IL-22 with the MAPK-ERK1/2 inhibitor PD98059 or the JAK/STAT inhibitor AG490 for 24 hours. Those HaCaT cells treated with phosphate buffered saline served as the control group. Subsequently, total proteins and mRNAs were extracted from the HaCaT cells. An immunofluorescence assay, Western blot and enzyme-linked immunosorbent assay（ELISA） were performed to determine the protein expression level of TIG3, and real-time fluorescence-based quantitative PCR to quantify the mRNA expression of TIG3 in HaCaT cells. Results The immunofluorescence assay showed that TIG3 protein was mainly expressed in the cytoplasm of HaCaT cells. As Western blot revealed, the protein expression level of TIG3 was 0.743 ± 0.035, 0.678 ± 0.040, 0.582 ± 0.041 and 0.328 ± 0.032 in HaCaT cells treated with IL-22 of 12.5, 25, 50 and 100 μg/L, respectively, significantly lower than that in the control group （0.839 ± 0.045, all P < 0.05）. ELISA also showed a decrease in the protein expression of TIG3 in IL-22-treated HaCaT cells, which was consistent with Western blot results. Further more, the mRNA expression level （2－△△Ct） of TIG3 was significantly weaker in HaCaT cells treated with IL-22 of 12.5, 25, 50 and 100 μg/L than in the control group （0.838 ± 0.036, 0.686 ± 0.061, 0.565 ± 0.047 and 0.457 ± 0.033 vs. 1.000, all P < 0.05）. The decrease in TIG3 mRNA and protein expressions was significantly attenuated in HaCaT cells treated with the combination of 50 μg/L IL-22 with PD98059 or AG490 compared with those treated with 50 μg/L IL-22 alone. Conclusion IL-22 can dose-dependently inhibit the expression of TIG3 in HaCaT cells, likely through the MAPK-ERK1/2 and JAK2/STAT3 signaling pathways.