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肿瘤坏死因子对人脐静脉内皮细胞PAI-1活性、mRNA的影响以及促分裂原活化蛋白激酶的作用
INFLUENCE OF TUMOR NECROSIS FACTOR α ON PAI-1 ACTIVITY AND mRNA LEVEL IN HUMAN UMBILICAL VEIN ENDOTHELIAL CELL AND THE MAPK REGULATORY EFFECT
【摘要】 目的 研究肿瘤坏死因子 α(TNF α)对人脐静脉内皮细胞 (HUVECs)纤溶酶原激活物抑制剂 (PAI 1)活性及mRNA水平的影响 ,以及促分裂原活化蛋白激酶 (MAPK)在其中的作用。 方法 进行HUVECs的培养和鉴定。加入不同浓度TNF α ,并选择最佳时间条件。采用发色底物法测定PAI 1活性 ,Northernblot法检测PAI 1mRNA的水平。运用MAPK激酶抑制剂PD980 5 9观察对上述PAI 1l表达系统的作用。 结果 不同浓度TNF α(5× 10 4IU L、1× 10 5IU L、2× 10 5IU L和 4× 10 5IU L)均明显增高HUVECsPAI 1的活性 (P <0 0 1) ,1× 10 5IU LTNF α作用不同时间 (12、18、2 4h) ,PAI 1活性均明显增加 (P <0 0 1)。 1× 10 5IU LTNF α作用 18h时 ,PAI 1mRNA水平为正常对照组的 2 88倍。MAPK激酶抑制剂PD980 5 9能显著抑制TNF α对PAI 1活性和mRNA表达增强的作用。 结论 TNF α增强HUVECsPAI 1活性与mRNA表达 ;PAI 1活性提高与其mRNA表达增加呈正相关 ;MAPK途径在TNF α诱导PAI 1反应中起着重要作用。
【Abstract】 Objective To study the influence of tumor necrosis factor α(TNF-α) on plasminogen activator inhibitor-1(PAI-1) activity and gene expression in human umbilical vein endothelial cell(HUVECs) and regulation of MAPK. Methods HUVECs were isolated,cultured and identified,The PAI-1 activity in HUVECs culture medium was measured by chromogenic assay.The concentration and time of TNF-α used to treat HUVECs were selected.The PAI-1 mRNA expression was determined by Northern blot.The effect of TNF-α on PAI-1 expression in HUVECs treated by MAPK inhibitor was observed. Results The PAI-1 activity was increased significantly when HUVECs were exposed to different concentrations and time of TNF-α in HUVECs.The PAI-1 mRNA level was increased in HUVECs after treatment with 1×10+5IU/L TNF-α for 18*!h.MAPK inhibitor(PD98059) could markedly inhibit the increase of PAI-1 activity and mRNA level induced by PAI-1.Conclusion The PAI-1 activity of HUVECs was increased after TNF-α exposure;The increase of PAI-1 activity was related to the increase of PAI-1 transcription; The MAPK may be involved in this process.
【Key words】 Tumor necrosis factor α(TNF-α); Plasminogen activator inhibitor-1(PAI-1); Human umbilical vein endothelial cell(HUVECs); Gene expression; Mitogen-activated protein kinase(MAPK);
- 【文献出处】 解剖学报 ,Acta Anatomica Sinica , 编辑部邮箱 ,2004年01期
- 【分类号】R363
- 【被引频次】2
- 【下载频次】124