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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。
Objective Previous studies have confirmed that tetramethylpyrazine has inhibitory effect on thrombin-induced tissue factor expression in human umbilical vein endothelial cells. This experiment further explored the mechanism of action of nitric oxide and nuclear factor kappa B pathway. Methods Human umbilical vein endothelial cells were cultured in RPMI-1640 complete medium; total coagulation activity was measured by one-phase coagulation; tissue factor mRNA was detected by reverse transcription polymerase chain reaction; immunohistochemical staining was used to study nuclear factor kappa B The shift. Results Nitric oxide synthase pathway blocker nitro-L-arginine methyl ethyl alone and human umbilical vein endothelial cell cells had no significant effect on the expression of tissue factor mRNA and total procoagulant activity (P>0.05). When nitro-L-arginine methyl ethyl ester, tetramethylpyrazine and thrombin were incubated together, the inhibitory effect of tetramethylpyrazine on thrombin-induced tissue factor expression in human umbilical vein endothelial cells was abolished (P<0.05). Immunohistochemical staining showed that the human umbilical vein endothelial cells were treated with thrombin for 45 min. The nucleus was brownishly stained. However, human umbilical vein endothelial cells were treated with tetramethylpyrazine for 15 min, and thrombin was added for 45 min. Coloring is significantly reduced. Conclusion The nitric oxide pathway is involved in the inhibitory effect of tetramethylpyrazine on thrombin-induced expression of tissue factor in vascular endothelial cells; Ligustrazine can inhibit the expression of tissue factor by affecting the activation of nuclear factor κB.