川芎嗪抗血栓形成的机制研究

Study on Mechanism of Ligustrazini Against Thrombosis

纤溶酶原激活物抑制剂-1是一种重要的促凝血因子。内毒素脂多糖通过刺激血管内皮细胞分泌纤溶酶原激活物抑制剂-1导致炎症过程中血栓形成。为探讨其机制，本文采用酶联免疫吸附试验法、North-ern印迹法和电泳迁移法观察中药川芎嗪对内毒素脂多糖所致内皮细胞纤溶酶原激活物抑制剂-1表达的影响及可能机制进行探讨。结果发现．川芎嗪不仅抑制内毒素脂多糖所致内皮细胞纤溶酶原激活物抑制剂-1蛋白分泌和mRNA表达．而且抑制内皮细胞纤溶酶原激活物抑制剂-1基础水平的表达。此结果提示，川芎嗪并非通过核因子NF－KB途径抑制内毒素脂多糖所致内皮细胞纤溶酶原激活物抑制剂-1表达增强的作用。

Aim To understand the mechanism of Chinese medicine ligustrazini against thrombosis and the effects of ligustrazini on plasminogen activator inhibitor type-1(PAI-1)expression in normal endothelial cells(EC)and EC exposed to lipopolysaccharide(LPS).Methods Human umbilical umbilical vein endothelial cells (HUVEC)were cultured by trypsin digestion method.PAI-1 protein in HUVEC conditioned medium was measured by sandwich enzyme-linked immunosorbent assay(ELISA),and PAI-1 mRNA expression was de-termined by Northern blot analysis.HUVEC nuclear factor-kappa B(NF-kB)nuclear translocation was as-sessed by electrophoretic mobility shift assay (EMSA).Results LPS treatment of cultured HUVEC result-ed in a significant increase in PAI-1 protein and mRNA expression by these cells.However,when HUVEC were incubated with LPS plus ligustrazini,the upregu-lation of PAI-1 by LPS was abated.Moreover ligus-trazini was able to decrease the basal level of PAI-1protein and mRNA as compared to control.Nuclear extracts prepared from LPS stimulated stimulated HUVEC demonstrated increased binding to the NF-kB oligonu-cleotide as compared to unstimulated cells,but ligus-trazini did not change those binding in the absence or presence of LPS.Conclusion Ligustrazini inhibited both basal and LPS-induced PAI-1 protein and mRNA expression in EC,and the modulation of PAI-1 in HUVEC by ligus-trazini might have other mechanisms rather than NF-kB pathway.