【摘要】 通过研究山莨菪碱对内毒素脂多糖致血管内皮细胞组织因子表达的影响 ,探讨山莨菪碱抗血栓形成以及治疗感染性休克的机制。采用酶消化法培养人脐静脉内皮细胞 ;用一步凝固法检测培养的内皮细胞组织因子活性 ;Northernblot检测内皮细胞组织因子mRNA的表达 ;为了评估上述作用是否通过核因子κB途径转导 ,采用电泳迁移变动检测法检测培养的内皮细胞核提取物核因子κBDNA结合活性。结果发现 ,内毒素脂多糖使内皮细胞组织因子活性及其mRNA表达显著增强 ;加入山莨菪碱后 ,内毒素脂多糖的这种作用明显减弱 ,并与山莨菪碱呈量效关系。且山莨菪碱能完全阻止内毒素脂多糖致内皮细胞核提取物核因子κBDNA结合活性。结果提示 ,山莨菪碱治疗感染性休克的机制之一是通过拮抗内毒素脂多糖致血管内皮细胞组织因子的表达 ,且这种拮抗作用可能通过、至少部分通过核因子κB途径转导更多还原

【Abstract】 Aim To study the effect of anisodamine on lipopolysaccharide (LPS) induced expression of tissue factor (TF) in vascular endothelial cells (EC). Methods Human umbilical vein endothelial cells (hUVEC) were cultured by trypsin digestion method. TF activity was measured in the lysates of hUVEC by using a single step clotting assay. Specific mRNA expressions were determined by Northern blotting. In order to evaluate a possible contribution of the nuclear factor κB (NF κB) pathway on anisodamine effects, electrophoretic mobility shift assays (EMSA) were performed using nuclear extracts from hUVEC and NF κB binding oligonucleotides. Results Treatment of hUVEC with LPS resulted in a significant increase in TF activity. Anisodamine dose dependently inhibited LPS induced upregulation of TF. These effects were also confirmed on the level of specific TF mRNA expression by Northern blotting. Furthermore, anisodamine completely abolished LPS induced NF κB DNA binding activity in nuclear extracts from hUVEC treated with LPS together with anisodamine. Conclusions Anisodamine counteracts endothelial cell activation by inhibiting LPS induced TF expression in these cells. Its interference with the NF κB pathway might at least partly contribute to this effect. The ability of anisodamine to counteract the effect of LPS on endothelial cells might be one underlying mechanism explaining its antithrombosis and efficacy in the treatment of bacteraemic shock.更多还原