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《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:

2016年第1期

页码:

16-019

栏目:

基础研究

出版日期:

2015-09-15

文章信息/Info

Title:

Effect of TNF-α on expression and activity of thrombomodulin and its mechanism

作者:

赖 斌¹; 魏玉杰¹; 刘惠亮¹; 李 屹¹; 谢玉凤¹; 付 玲²

（1.武警总医院心血管内科，北京100039；
2.军事医学科学院微生物流行病学研究所，北京 100071）

Author(s):

LAI Bin¹;  WEI Yu-jie¹;  LIU Hui-liang¹;  LI Yi¹;  XIE Yu-feng¹;  FU Ling²

(1.Department of Cardiology, General Hospital, Chinese People's Armed Police Forces, Beijing 100039, China；
2.Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China)

关键词:

肿瘤坏死因子-α; 血栓调节蛋白; NF-κB; 脐静脉内皮细胞

Keywords:

tumor necrosis factor-α;  thrombomodulin;  NF-κB;  human umbilical vein endothelial cells

分类号:

R392.12

DOI:

-

文献标识码:

A

摘要:

目的 观察肿瘤坏死因子(TNF)-α对血栓调节蛋白（TM）在原代人脐静脉内皮细胞（HUVECs）的表达和活性的影响，并初步探讨其作用机制。方法 体外培养HUVECs，实验分为3组：①对照组：加入与TNF-α等体积的PBS培养细胞6 h；②TNF-α组：TNF-α终浓度为10 ng/ml孵育细胞6 h；③BAY11-7082抑制组：BAY11-7082终浓度2 μg/ml孵育细胞1 h阻断NF-κB通路后再用10 ng/ml的TNF-α干预细胞6 h。分别采用流式细胞技术、荧光定量聚合酶链反应（RT-PCR）和酶标仪检测TM蛋白、mRNA表达及活性强度。结果 TNF-α组相比与对照组和BAY11-7082抑制组在TM的蛋白、mRNA水平上均明显降低（P<0.05），TM活性也明显减弱(P<0.05)。而BAY11-7082抑制组相比与对照组在TM蛋白水平和活性均无差异，在mRNA水平上升高(P<0.05)。结论 TNF-α可明显降低TM表达，并抑制其活性，NF-κB通路参与了这一过程。

Abstract:

AIM To observe the effect of tumor necrosis factor-α (TNF-α) on the expression and activity of thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs) and to investigate its mechanism. METHODSHUVECs were incubated in vitro and the experiment was conducted using three groups: control group, TNF-α group (incubated with TNF-α 10 ng/ml for 6 h) and BAY11-7082 suppression group ［first incubated with BAY11-7082 2 μg/ml for 1h, then incubated with TNF-α (10 ng/ml) for 6 h］. Flow cytometric and RT-PCR technique were used to detect TM protein and mRNA, and Microplate Reader was used to detect TM activity. RESULTSCompared with those in the control group and BAY11-7082 suppression group, both mRNA and protein levels of TM were significantly reduced (P<0.05) and the TM activity in HUVECs was inhibited (P<0.05) in TNF-α group. However, compared with those in the control group, no statistical differences were observed in the protein level of TM and the TM activity, whereas mRNA levels of TM were increased (P<0.05) in the BAY11-7082 suppression group. CONCLUSIONExposure to TNF-α significantly decreases the expression of TM and inhibits the TM activity in HUVECs. NF-κB pathway plays a role in this process.

参考文献/References

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备注/Memo

备注/Memo:

收稿日期：2014-10-18.
>基金项目：国家自然科学基金项目资助（81370315）
通讯作者：刘惠亮，教授，主要从事心血管病介入治疗研究 Email:huiliangl531@163.com
作者简介：赖斌，硕士生 Email: laibintiantian@163.com

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更新日期/Last Update: 2015-09-16