中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (4): 256-259.doi: 10.3760/cma.j.issn.0412-4030.2018.04.003

• 论著 • 上一篇    下一篇

MicroRNA-143对白介素13诱导的人角质形成细胞组织激肽释放酶7表达的影响

曾跃平1,朱晨雨2,陈程1,贾倩楠3,晋红中4   

  1. 1. 中国医学科学院北京协和医学院北京协和医院
    2. 北京协和医院
    3. 中国医学科学院 北京协和医院
    4. 中国医学科学院北京协和医学院北京协和医院皮肤科
  • 收稿日期:2017-05-02 修回日期:2017-06-16 发布日期:2018-03-29
  • 通讯作者: 晋红中 E-mail:jinhongzhong@263.net
  • 基金资助:
    国家自然科学基金;北京市自然科学基金;2015年度北京市优秀人才青年骨干个人项目;北京协和医院中青年科研基金

Effect of microRNA-143 on interleukin-13-induced of kallikrein 7 in human epidermal keratinocytes

Yue-ping ZENG1, 2,Cheng Chen1, 3,   

  • Received:2017-05-02 Revised:2017-06-16 Published:2018-03-29
  • Supported by:
    National Natural Science Foundation of China;Beijing Municipal Natural Science Foundation;Training Program Foundation for the Excellent Youth Scholars of Beijing in 2015;Young Scientific Research Fund of Peking Union Medical College Hospital

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摘要： 目的 探讨microRNA?143（miR?143）对白介素13（IL?13）诱导的人角质形成细胞组织激肽释放酶7（KLK7）表达的影响。方法 取对数生长期人皮肤原代角质形成细胞（NHEK），分别用0、2、10、50 μg/L IL?13处理24 h，或用50 μg/L IL?13分别处理0、6、12、24、48 h后，收集细胞，提取细胞总RNA，实时荧光定量PCR检测KLK7基因mRNA水平。再取部分NHEK，分为4组，即NHEK组（空白对照组，既不转染也不用IL?13刺激）、IL?13组（仅用50 μg/L IL?13处理）、miR?NC组（转染microRNA mimics阴性对照后用50 μg/L IL?13处理）和miR?143组（转染miR?143 mimics后用50 μg/L IL?13处理），IL?13处理24 h后，实时荧光定量PCR检测各组中KLK7 mRNA的相对表达水平，Western免疫印迹检测KLK7蛋白表达水平。结果 0、2、10、50 μg/L IL?13处理NHEK 24 h后，KLK7 mRNA的相对表达水平分别为1.00 ± 0.12、0.89 ± 0.04、1.15 ± 0.09和1.70 ± 0.10，随着IL?13浓度的升高，KLK7 mRNA相对表达水平有上升趋势（F = 92.48，P＜0.05）。50 μg/L IL?13分别处理NHEK 0、6、12、24、48 h后，KLK7 mRNA相对表达水平分别为1.00 ± 0.05、1.05 ± 0.12、1.71 ± 0.20、1.97 ± 0.19和2.48 ± 0.13，随着IL?13处理时间延长，KLK7 mRNA的相对表达水平有上升趋势（F = 206.44，P＜0.05）。与miR?NC组KLK7 的mRNA和蛋白相对表达水平相比，miR?143组均降低，差异有统计学意义（t值分别为6.76、4.23，均P＜0.05）。结论 在人角质形成细胞中，IL?13上调KLK7的表达可能与miR?143的调控相关。

关键词: 皮炎, 特应性, 角质形成细胞, 白细胞介素13, 组织激肽释放酶7, 微RNA-143

Abstract: Zeng Yueping, Zhu Chenyu, Chen Cheng, Jia Qiannan, Jin Hongzhong Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Center for Translational Medicine, Beijing 100730, China Corresponding author: Jin Hongzhong, Email: jinhongzhong@263.net 【Abstract】 Objective To evaluate the effect of microRNA-143 （miR-143） on interleukin （IL）-13-induced of kallikrein 7（KLK7） in primary normal human epidermal keratinocytes（NHEKs）. Methods Some NHEKs at exponential growth phase were divided into 4 groups to be treated with recombinant human IL-13 at different concentrations of 0, 2, 10 and 50 μg/L respectively for 24 hours, and some NHEKs were treated with 50 μg/L IL-13 for 0, 6, 12, 24 and 48 hours separately. After the treatment, NHEKs were collected, and total RNA was extracted. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA of KLK7. Some other NHEKs were divided into another 4 groups: NHEK group （blank control group） receiving no treatment, IL-13 group treated with 50 μg/L IL-13, miR-NC group transfected with miRNA mimics negative control followed by the treatment with 50 μg/L IL-13, and miR-143 group transfected with miR-143 mimics followed by the treatment with 50 μg/L IL-13. After 24-hour treatment with IL-13, real-time fluorescence-based quantitative PCR and Western blot analysis were conducted to determine the mRNA and protein of KLK7 respectively in the above groups. Results After 24-hour treatment with IL-13 at concentrations of 0, 2, 10 and 50 μg/L, the mRNA of KLK7 in NHEKs was 1.00 ± 0.12, 0.89 ± 0.04, 1.15 ± 0.09 and 1.70 ± 0.10 respectively, and significantly increased along with the increase of IL-13 concentrations （F = 92.48, P < 0.05）. After 0-, 6-, 12-, 24- and 48-hour treatment with 50 μg/L IL-13, the mRNA of KLK7 in NHEKs was 1.00 ± 0.05, 1.05 ± 0.12, 1.71 ± 0.20, 1.97 ± 0.19 and 2.48 ± 0.13 respectively, and significantly increased over time （F = 206.44, P < 0.05）. Compared with the miR-NC group, the miR-143 group showed significantly decreased mRNA and protein of KLK7 （t = 6.76, 4.23 respectively, both P < 0.05）. Conclusion In NHEKs, IL-13 can up-regulate the of KLK7, likely by the regulation of miR-143.

Key words: Dermatitis, atopic, Keratinocytes, Interleukin-13, Kallikrein 7, MicroRNA-143

引用本文

曾跃平 朱晨雨 陈程 贾倩楠 晋红中. MicroRNA-143对白介素13诱导的人角质形成细胞组织激肽释放酶7表达的影响[J]. 中华皮肤科杂志, 2018,51(4):256-259. doi:10.3760/cma.j.issn.0412-4030.2018.04.003

Yue-ping ZENG Cheng Chen. Effect of microRNA-143 on interleukin-13-induced of kallikrein 7 in human epidermal keratinocytes[J]. Chinese Journal of Dermatology, 2018, 51(4): 256-259.doi:10.3760/cma.j.issn.0412-4030.2018.04.003