天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (4): 384-389.doi: 10.11958/20202041

• 实验研究 • 上一篇    下一篇

CD137-CD137L信号通路通过活化TNF受体相关因子6促进小鼠动脉粥样硬化斑块内血管新生#br#

翁嘉懿1，殷云杰2，马雪兴1，李渊1，陈璐1，何洪涛1，徐桂冬1△，孙康云1   

  1. 1南京医科大学附属苏州医院心血管内科（邮编215000）；2宜兴市人民医院心血管内科
  • 收稿日期:2020-07-17 修回日期:2020-12-21 出版日期:2021-04-15 发布日期:2021-04-16
  • 通讯作者: 徐桂冬 E-mail:guidong_xu@126.com
  • 作者简介:翁嘉懿（1991），女，硕士，主治医师，主要从事动脉粥样硬化发病机制及治疗研究。E-mail：wengjiayi129@126.com
  • 基金资助:
    苏州市“科教兴卫”青年科技项目（KJXW2017040）；苏州市科技局项目（sys2018088）

CD137-CD137L signaling promotes angiogenesis in atherosclerosis plaque of mice through activating TNF receptor associated factor 6

WENG Jia-yi1，YIN Yun-jie2， MA Xue-xing1,  LI Yuan1, CHEN Lu1, HE Hong-tao1， XU Gui-dong1△， SUN Kang-yun1   

  1. 1 Department of Cardiovascular Diseases, Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou 215000, China;     2 Department of Cardiovascular Diseases, Yixing People’s Hospital
  • Received:2020-07-17 Revised:2020-12-21 Published:2021-04-15 Online:2021-04-16
  • Contact: XU Gui-dong E-mail:guidong_xu@126.com

PDF (PC)

4318

摘要： 目的　探讨CD137-CD137L信号通路是否通过活化TNF受体相关因子6（TRAF6）促进粥样硬化斑块内血管新生。方法　将小鼠内皮细胞（bEnd.3）和小鼠主动脉环分别分为对照组（培养基中加入10 μg/L TNF-α）、IgG同型对照组（培养基中加入10 μg/L TNF-α+5 mg/L IgG2b）和CD137刺激组（培养基中加入10 μg/L TNF-α+5 mg/L CD137抗体）。利用转染小干扰RNA（siRNA）技术抑制内皮细胞和主动脉环TRAF6基因表达，将内皮细胞和主动脉环分别分为对照siRNA组（转染对照siRNA）和TRAF6 siRNA组（转染TRAF6 siRNA）。分别采用Western blot和实时荧光定量PCR（qPCR）检测内皮细胞和主动脉环TRAF6、血管内皮生长因子（VEGF）、Smad1/5蛋白和mRNA的表达；采用Transwell迁移实验检测内皮细胞的迁移能力；内皮细胞管腔形成实验检测内皮细胞的管腔形成能力；主动脉环血管新生实验检测主动脉环新生微血管形成能力。结果　CD137刺激组内皮细胞和主动脉环TRAF6、VEGF蛋白和mRNA表达水平均高于对照组和IgG同型对照组（均P＜0.05）。与对照siRNA组比较，TRAF6 siRNA组内皮细胞和主动脉环TRAF6、Smad1/5蛋白和mRNA表达水平明显降低，内皮细胞迁移数量减少，内皮细胞小管长度和分支数量降低，主动脉环新生微血管数量减少（均P＜0.05）。结论　CD137-CD137L信号通路可能通过TRAF6促进小鼠动脉粥样硬化斑块内血管新生。

关键词: 抗原, CD137, 动脉粥样硬化, TNF受体相关因子6, Smad1蛋白质, Smad5蛋白质, 血管新生

Abstract: Objective　To explore whether CD137-CD137L signaling pathway can promote angiogenesis in atherosclerosis plaque via activating TNF receptor associated factor 6 (TRAF6). Methods　Endothelial cells (bEnd.3) and the aortic rings from male mice were divided into the following groups: control group (with 10 μg/L TNF-α in culture medium), IgG isotype control group (with 10 μg/L TNF-α+5 mg/L IgG2b in culture medium) and CD137 activated group (with 10 μg/L TNF-α+5 mg/L CD137 antibody in culture medium). Small interfering RNA (siRNA) oligonucleotide was used for TRAF6 gene knockdown in MBVEC and the aortic rings, which were divided into two groups: control siRNA group (control siRNA transfected) and TRAF6 siRNA group (TRAF6 siRNA transfected). Western blot assay was used to determine protein expressions of TRAF6, Smad1/5 and VEGF. Transwell assay was used to observe the migration ability of endothelial cells. Matrigel tube formation was used to test the tube formation ability of endothelial cells and the mouse aortic rings. Angiogenesis assay were used for detecting the aortic ring microangiogenesis. Results　The expression levels of mRNA and protein of TRAF6 and VEGF were significantly higher in CD137 activated group than those in IgG isotype control group and  control group (both in the endothelial cells and aortic rings, P＜0.05). The expression levels of mRNA and protein of TRAF6 and Smad1/5 were significantly lower in TRAF6 siRNA group than those in control siRNA group in endothelial cells and aortic rings. Migration cell number was remarkably lower in TRAF6 siRNA group than that in control siRNA group. Values of the formation of the tube lengthand branch number were both significantly lower in TRAF6 siRNA group than those in control siRNA group. The numbers of microvessels outgrowth were dramatically reduced in aortic rings in control siRNA group (P＜0.05). Conclusion　CD137-CD137L signaling pathway can promote angiogenesis in atherosclerosis plaque by activating TRAF6.

Key words: antigens, CD137, atherosclerosis, TNF receptor-associated factor 6, Smad1 protein, Smad5 protein, angiogenesis

中图分类号: