川芎嗪抑制凝血酶诱导血管内皮细胞组织因子表达的机制
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湖南省教育厅科研基金资助项目(06C783);;湘南学院科研基金资助项目(05Y035)


Mechanisms for Tetramethylpyrazine on the Expression of Tissue Factor Induced by Thrombin in Vascular Endothelial Cells
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    摘要:

    目的以前的研究已证实川芎嗪对凝血酶诱导人脐静脉内皮细胞株表达组织因子具有抑制效应。本实验进一步探讨一氧化氮及核因子κB途径在其中所发挥的作用机制。方法人脐静脉内皮细胞细胞培养采用RPMI-1640完全培养基;一期凝固法测总促凝活性;组织因子mRNA采用逆转录聚合酶链式反应方法检测;免疫组织化学染色用于研究核因子κB的移位。结果一氧化氮合酶途径阻断剂硝基左旋精氨酸甲基乙酯单独和人脐静脉内皮细胞细胞孵育时对细胞表达组织因子mRNA和总促凝活性没有明显的影响(P>0.05);硝基左旋精氨酸甲基乙酯、川芎嗪和凝血酶三者共同孵育时,川芎嗪抑制凝血酶诱导人脐静脉内皮细胞细胞组织因子表达的作用被取消(P<0.05)。免疫组织化学染色显示人脐静脉内皮细胞细胞经凝血酶处理45min后,细胞核内棕黄色着色明显,但人脐静脉内皮细胞细胞经川芎嗪处理15min后,再加入凝血酶作用45min,核内棕黄色着色则明显减少。结论一氧化氮途径参与了川芎嗪抑制凝血酶诱导血管内皮细胞表达组织因子的作用;川芎嗪能够通过影响核因子κB的活化来抑制组织因子的表达。

    Abstract:

    Aim Previous study demonstrated that tetramethylpyrazine(TMP)had an inhibitory effect on the TF expression induced by thrombin in human umbilical vein endothelium derived cell line human umbilical vascular endothelial cells(HUVEC).However,its mechanisms were not fully understood.In this study,we observed the role of nitric oxide(NO)and transcription factor NF-κB in the regulation of TMP on tissue factor(TF)expression.Methods HUVEC were cultured in RPMI-1640.TF activity was determined with one-stage clotting assay measuring total cellular pro-coagulant activity(PCA).TF mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).Immunohistochemical analysis was performed to evaluate the translocation of NF-κB.Results NG-nitro-L-arginine methylate(L-NAME)is an inhibitor of nitric oxide synthase(NOS),and it alone had no marked effects on PCA and TF mRNA in HUVEC,but L-NAME significantly decrease the inhibitory effects of TMP on PCA increment induced by thrombin(P<0.05).Immunohistochemical analysis demonstrated that NF-κB translocation from cytoplasm to the nucleus was found after treatment of endothelial cells with thrombin.TMP inhibited thrombin-induced NF-κB translocation from cytoplasm to the nucleus.Conclusion NO pathway participates in the inhibitory effect of TMP on the expression of TF induced by thrombin.TMP can inhibit TF expression induced by thrombin through suppressing NF-κB activation.

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成春英,孙勇,文志斌,何晓凡,王谷丰,林国强,蒋海河,唐先明,贺石林.川芎嗪抑制凝血酶诱导血管内皮细胞组织因子表达的机制[J].中国动脉硬化杂志,2010,18(3):184~188.

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  • 收稿日期:2009-11-25
  • 最后修改日期:2010-01-12
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