Chinese Medical Sciences Journal ›› 2019, Vol. 34 ›› Issue (3): 199-204.doi: 10.24920/003489

• 论著 • 上一篇    下一篇

白细胞介素-36β对体外培养角质形成细胞炎症反应与分化的影响

王文明1,吴超1,余晓玲2,晋红中1,*()   

  1. 1 中国医学科学院 北京协和医学院 北京协和医院皮肤科,北京 100730
    2 南方医科大学皮肤病医院皮肤科,广州 510000
  • 收稿日期:2018-12-28 出版日期:2019-09-30 发布日期:2019-10-11
  • 通讯作者: 晋红中 E-mail:jinhongzhong@263.net

IL-36β Promotes Inflammatory Activity and Inhibits Differentiation of Keratinocytes In Vitro

Wang Wenming1,Wu Chao1,Yu Xiaoling2,Jin Hongzhong1,*()   

  1. 1 Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China
    2 Department of Dermatology, Dermatology Hospital of Southern Medical University, Guangdong Provincial Dermatology Hospital, Guangdong 510080, China
  • Received:2018-12-28 Published:2019-09-30 Online:2019-10-11
  • Contact: Jin Hongzhong E-mail:jinhongzhong@263.net

摘要:

目的 银屑病是一种免疫介导的炎症性疾病。尽管目前银屑病的研究取得了巨大的进展,但其确切机制尚不完全清楚。表皮的增殖异常在银屑病的发病中发挥重要作用。本研究将在体外探究白细胞介素-36β(IL-36β)对角质形成细胞功能的影响。

方法 体外培养人永生化角质形成细胞系HaCaT,分为3组:对照组、50 ng/ml IL-36β组、100 ng/ml IL-36β组,分别处理24 h。采用CCK8法检测细胞增殖,流式细胞术检测细胞凋亡与细胞周期分布,实时荧光定量PCR检测角质形成细胞分化指标keratin 10 mRNA和involucrin mRNA的表达,ELISA法检测培养液中炎症因子IL-1β和IL-6的含量。

结果 与对照组相比,50、100 ng/ml IL-36β可使HaCaT细胞阻滞在S期(P<0.05);同时可抑制细胞keratin 10、involucrin mRNA的表达(P<0.01);100 ng/ml IL-36β可增加细胞培养液中IL-1β与IL-6的含量(P<0.05)。

结论 IL-36β可能导致角质形成细胞周期阻滞,抑制角质形成细胞分化,促进角质形成细胞的炎症反应。

关键词: 白细胞介素-36β;, 银屑病, 角质形成细胞, 炎症反应, 细胞分化

Abstract:

Objective Psoriasis is an immune-mediated inflammatory disease. Despite advances in the study of its pathogenesis, the exact development mechanism of psoriasis remains to be fully elucidated. Hyperproliferative epidermis plays a crucial role in psoriasis. This study aimed to investigate the effects of interleukin-36β (IL-36β) on keratinocyte dysfunction in vitro.

Methods Human keratinocyte cell lines, HaCaT cells, were treated with 0 (control), 50 or 100 ng/ml IL-36β respectively for 24 h. Cell viability was determined with a cell counting kit-8 assay. Flow cytometry was used to assess the effects of IL-36β on apoptosis and cell cycle distribution. Expressions of the differentiation markers, such as keratin 10 and involucrin, were evaluated by quantitative real-time polymerase chain reaction (RT-qPCR). Expressions of the inflammatory cytokines, IL-1β and IL-6 were tested by ELISA.

Results CCK8 assay showed the survival rate had no significant difference between the control and treated group (P > 0.05). Flow cytometry analysis showed cell cycle arrest at S phase in the IL-36β-treated groups compared with the control group (P < 0.05). RT-qPCR verified the decreased mRNA expressions of keratin 10 and involucrin in the IL-36β-treated groups compared with the negative control (P < 0.01). ELISA showed 100 ng/ml IL-36β enhanced levels of IL-1β and IL-6 in culture supernatants of HaCaT cells compared with the negative control (P < 0.05).

Conclusion Taken together, these findings suggest that IL-36β could induce cell cycle arrest at S phase, inhibit keratin 10 and involucrin expressions and promote inflammatory activity in HaCaT cell lines.

Key words: interleukin-36β;, psoriasis, keratinocytes, inflammatory activity, differentiation

Copyright © 2018 Chinese Academy of Medical Sciences. All right reserved.
 
www.cmsj.cams.cn
京公安备110402430088 京ICP备06002729号-1  Powered by Magtech.

Supervised by National Health & Family Plan Commission of PRC

9 Dongdan Santiao, Dongcheng district, Beijing, 100730 China

Tel: 86-10-65105897  Fax:86-10-65133074 

E-mail: cmsj@cams.cn  www.cmsj.cams.cn

Copyright © 2018 Chinese Academy of Medical Sciences

All right reserved.

京公安备110402430088  京ICP备06002729号-1