天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (12): 1233-1236.doi: 10.11958/20170738

• 细胞与分子生物学 •    下一篇

microRNA-145 通过 CD40 对泡沫细胞 免疫炎症反应的影响

刘恩照△， 路利平， 刘运龄， 梁雪， 李广平   

  1. 作者单位： 天津医科大学第二医院心脏科， 天津市心血管病离子与分子机能重点实验室， 天津心脏病学研究所 （邮编 300211） 作者简介： 刘恩照 （1979）， 男， 副主任医师， 主要从事心律失常与冠心病基础与临床研究 △通讯作者 E-mail：liu_ezh@126.com
  • 收稿日期:2017-06-26 修回日期:2017-09-19 出版日期:2017-12-15 发布日期:2017-12-15
  • 通讯作者: 刘恩照 E-mail:liu_ezh@126.com

The effect of microRNA-145 on immune-inflammatory response of foam cells by targeting CD40

LIU En-zhao△, LU Li-ping, LIU Yun-ling, LIANG Xue, LI Guang-ping   

  1. Department of Cardiology, the Second Hospital of Tianjin Medical University, Tianjin Key Laboratory of Ionic-Molecular Function of Cardiovascular Disease, Tianjin Institute of Cardiology, Tianjin 300211, China △Corresponding Author E-mail: liu_ezh@126.com
  • Received:2017-06-26 Revised:2017-09-19 Published:2017-12-15 Online:2017-12-15
  • Contact: Enzhao LIU E-mail:liu_ezh@126.com

PDF (PC)

3583

摘要： 目的 探讨 microRNA（miR） -145 通过 CD40 对泡沫细胞免疫炎症反应的影响。方法 体外培养小鼠巨 噬细胞系 RAW 264.7 细胞， 随机均分为模型组（不转染）、 miR-145 mimics 组（转染 miR-145 mimics）、 miR-145 inhibitor 组（转染 miR-145 inhibitor）、 沉默 CD40 序列组（转染 siCD40）， 转染 6 h 之后， 用氧化低密度脂蛋白（oxLDL）刺激 24 h 诱导形成泡沫细胞。应用 real-time qPCR（RT-qPCR）和 Western blot 检测各组细胞中 CD40 mRNA 和蛋白的表达情况； ELISA 法检测细胞上清液中炎症因子白细胞介素（IL） -1、 IL-6、 肿瘤坏死因子（TNF） -α 水平。 结果 与模型组相比， miR-145 mimics 组 CD40 mRNA、 蛋白及 IL-1、 IL-6、 TNF-α 的表达水平均明显下降（P < 0.01）； 经 miR-145 抑制剂干预后， 上述指标均较模型组和 miR-145 mimics 组升高（P < 0.01）； 而转染沉默 CD40 序 列后， CD40 mRNA、 蛋白及 IL-1、 IL-6、 TNF-α 的表达水平较 miR-145 抑制剂组均明显下降 （P < 0.01）。结论 miR- 145 可通过靶基因 CD40 调控泡沫细胞免疫炎症过程， 抑制 CD40/CD40L 信号通路活化， 抑制炎症反应。

关键词: 微 RNAs, 抗原, CD40, 泡沫细胞, 转染, 白细胞介素 6, 肿瘤坏死因子 α, miR-145 中图分类

Abstract: Objective To investigate the effect of miRNA-145 (miR-145) on immuno-inflammatory reaction of foam cells by targeting CD40. Methods Mouse macrophage cell line RAW 264.7 cells cultured in vitro were randomly divided into model group (non-transfected), miR-145 mimics group (transfected miR-145 mimics), miR-145 inhibitor group (transfected miR-145 inhibitor) and silencing CD40 sequence group (transfected siCD40). Then oxidized low density lipoprotein (ox-LDL) was used to stimulate for 24 h to establish immune inflammatory damage cell model. Quantitative realtime polymerase chain reaction (RT-qPCR) and Western blot assay were used to detect the levels of CD40 mRNA and protein of each group. ELISA was used to detect the levels of inflammatory factors interleukin (IL) - 1, IL-6 and tumor necrosis factor (TNF) - α in cell supernatant. Results Compared with model group, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-α were all significantly decreased in miR-145 mimics group (P < 0.01). After transfected with miR-145 inhibitor, the above indexes were all significantly increased than those of model group and miR-145 mimics group (P < 0.01). After transfected with CD40 siRNA, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-α were all obviously decreased compared with those of miR-145 inhibitor group (P < 0.01). Conclusion MiR-145 can regulate the immune inflammatory process of foam cells through the target gene CD40, inhibit the activation of CD40/CD40L signaling pathway and inhibit inflammatory response.

Key words: microRNAs, antigens, CD40, foam cells, transfection, interleukin-6, tumor necrosis factor-alpha, miR-145