| TNF-α对血栓调节蛋白表达活性的影响及其作用机制的探讨 |
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| 引用本文： | 赖斌,魏玉杰,刘惠亮,李屹,谢玉凤,付玲.TNF-α对血栓调节蛋白表达活性的影响及其作用机制的探讨[J].心脏杂志,2016,28(1):16-19. |
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| 作者姓名： | 赖斌 魏玉杰 刘惠亮 李屹 谢玉凤 付玲 |
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| 作者单位： | 1. 武警总医院心血管内科,北京,100039; |
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| 基金项目： | 国家自然科学基金项目资助(81370315) |
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| 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;2 TNF-α组:TNF-α终浓度为10 ng/ml孵育细胞6 h;3 BAY11-7082抑制组:BAY11-7082终浓度2μg/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。
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| 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 |
| 收稿时间： | 2014-10-18 |
Effect of TNF-α on expression and activity of thrombomodulin and its mechanism |
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| Affiliation: | 1. Department of Cardiology,General Hospital,Chinese People’s Armed Police Forces;2. Institute of Biotechnology,Academy of Military Medical Sciences |
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| 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. METHODS HUVECs 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. RESULTS Compared 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. CONCLUSION Exposure 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. |
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