中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (9): 636-640.

• 论著 • 上一篇    下一篇

表没食子儿茶素没食子酸酯抑制白细胞介素17诱导的角质形成细胞增殖和凋亡

付丹丹1,胡华2,2,孙敏2,李敏河南3,田中伟3   

  1. 1. 新乡医学院第一附属医院皮肤科
    2. 新乡医学院第一附属医院
    3. 河南省新乡医学院第一附属医院皮肤科
  • 收稿日期:2015-11-25 修回日期:2016-01-22 出版日期:2016-09-15 发布日期:2016-08-30
  • 通讯作者: 田中伟 E-mail:zhonwt@163.com
  • 基金资助:

    河南省教育厅科学技术研究重点项目;新乡市科技发展计划项目;.河南生卫生科技创新人才型工程

Epigallocatechin gallate inhibits the proliferation and apoptosis of keratinocytes induced by interleukin-17

  • Received:2015-11-25 Revised:2016-01-22 Online:2016-09-15 Published:2016-08-30

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摘要：

目的 探讨表没食子儿茶素没食子酸酯（EGCG）对白细胞介素17（IL?17）诱导的角质形成细胞损伤的保护作用及其机制。 方法 实验分空白对照组、IL?17组、IL?17 + EGCG组、IL?17 + SP600125组和IL?17 + EGCG + 茴香霉素组。CCK?8试剂盒检测细胞增殖；流式细胞仪检测细胞凋亡；ELISA试剂盒检测IL?6、IL?23和 IL?8的表达；Western 印迹检测JNK和P?JNK的表达。 结果 IL?17促进HaCaT细胞增殖，且增殖率与IL?17浓度有关，90 μg/L IL?17组的细胞增殖率最高（P < 0.05）。60 μmol/L EGCG显著抑制90 μg/L IL?17诱导的细胞增殖（P < 0.05），促进细胞凋亡（P < 0.05），降低IL?6、IL?23和 IL?8的表达（P < 0.05）。与IL?17组相比，IL?17 + EGCG组、IL?17 + SP600125组的P?JNK表达显著下调（P < 0.05），细胞增殖率降低（P < 0.05），IL?6、IL?23和 IL?8的表达减少（P < 0.05）；与IL?17 + EGCG组相比，IL?17 + EGCG + 茴香霉素组的P?JNK表达显著上调（P < 0.05），细胞增殖率和IL?6、IL?23、IL?8的表达均明显上升（P < 0.05）。 结论 EGCG对IL?17诱导的HaCaT细胞增殖凋亡、炎症反应等损伤具有保护作用，其保护作用可能与抑制JNK信号通路有关。

Abstract:

Fu Dandan, Hu Hua, Sun Min, Li Min, Tian Zhongwei Department of Dermatology and Venereology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan, China Corresponding author: Tian Zhongwei, Email: zhonwt@163.com 【Abstract】 Objective To evaluate the protective effect of epigallocatechin gallate （EGCG） against interleukin （IL）-17-induced injury to keratinocytes, and to explore its mechanism. Methods Some cultured HaCaT cells were divided into 3 groups to be treated with IL-17 alone at concentrations of 50, 70, 90 μg/L, respectively, with those receiving no treatment as the blank control group. Some HaCaT cells were divided into 5 groups: IL-17 group treated with 90 μg/L IL-17 alone, IL-17 + EGCG group treated with 90 μg/L IL-17 and 60 μmol/L EGCG, IL-17 + SP600125 group treated with 90 μg/L IL-17 and SP600125 （a JAK signaling pathway inhibitor）, IL-17+ EGCG + anisomycin group treated with 90 μg/L IL-17, 60 μmol/L EGCG and anisomycin （a Janus kinase signaling pathway activator）, and blank control group receiving no treatment. After different durations of treatment, CCK-8 assay was performed to evaluate cellular proliferative activity, flow cytometry to detect cell apoptosis, enzyme-linked immunosorbent assay （ELISA） to measure levels of IL-6, IL-23 and IL-8, and Western-blot analysis to determine protein s of c-Jun N-terminal kinase （JNK） and phosphorylated JNK （P-JNK）. Results IL-17 promoted cellular proliferation of HaCaT cells, and the proliferation rate, which was correlated with the concentration of IL-17, reached the maximum in the 90-μg/L IL-17 group （P < 0.05）. EGCG at 60 μmol/L significantly inhibited cellular proliferation of, promoted apoptosis in, and reduced IL-6, IL-23 and IL-8 s in, HaCaT cells induced by 90 μg/L IL-17 （all P < 0.05）. Compared with the IL-17 group, the IL-17 + EGCG group and IL-17 + SP600125 group both showed significantly decreased P-JNK , cell proliferation rate and IL-6, IL-23 and IL-8 levels （all P < 0.05）. However, compared with the IL-17 + EGCG group, the IL-17 + EGCG + anisomycin group showed significantly increased protein of P-JNK, cell proliferation rate and IL-6, IL-23 and IL-8 levels （all P < 0.05）. Conclusion EGCG protected against IL-17-induced injury to HaCaT cells, such as abnormal cell proliferation, apoptosis and inflammatory response, likely by inhibiting the JNK signaling pathway.