ASO血瘀证血清对血管内皮细胞损伤的影响及桃红四物汤的调节作用

Effect of ASO Blood Stasis Syndrome Serum on Vascular Endothelial Cell Injury and Regulation of Taohong Siwu Decoction on it

目的探讨动脉硬化闭塞症(arteriosclerosis obliterans,ASO)血瘀证血清对血管内皮细胞损伤的影响及桃红四物汤的调节作用。方法建立脐静脉内皮细胞培养体系,制成ASO血瘀证血清内皮细胞损伤模型,分别加入桃红四物汤低、中、高浓度大鼠含药血清,MTT法观察内皮细胞增殖活性,透射电镜观察内皮细胞超微结构,激光共聚焦显微镜观察细胞内钙离子浓度和细胞骨架的变化,酶联免疫吸附(E LISA)观察各组内皮细胞培养液上清中内皮素(ET)、一氧化氮(NO)和转化生长因子β_1(transforming growth factorβ_1,TGF-β_1)的含量。结果 ASO血瘀证血清组内皮细胞增殖活性下降,细胞结构破坏明显,钙离子浓度升高,培养液中ET、NO及TGF-β_1含量明显升高(P<0.01),ET/NO比例失衡。与桃红四物汤含药血清共孵育后,内皮细胞增殖活性和细胞结构的损伤明显改善,ET、NO及TGF-β_1水平下降(P<0.05,P<0.01),ET/NO比例趋于正常。结论桃红四物汤治疗血瘀证ASO的主要机制为改善血管内皮细胞的损伤和分泌功能失调。

Objective To explore the effect of arteriosclerosis obliterans（ASO）blood stasis syndrome（BSS）serum on vascular endothelial cell injury and to study the regulation of Taohong Siwu Decoction（TSD）on it.Methods Umbilical vein endothelial cell culture system was established.The serum endothelial cell injury model with ASO BSS was prepared.Low,medium,and high concentrations TSD containing serums were respectively added.The endothelial cell proliferation activity was observed by MTT method.Ultrastructures of endothelial cells were observed under transmission electron microscope.Changes of intracellular calcium ion concentration and the cytoskeleton were observed under laser confocal microscope.Contents of ET,NO,and transforming growth factor beta_1（TGF-β_1）in endothelial cell culture supernatant were detected by ELISA.Results In ASO BSS serum group endothelial cell proliferation activities decreased,the cell structure was obviously destroyed,calcium ion concentration increased,contents of ET,NO and TGF-β_1 increased significantly（P0.01）,and ET/NO ratio was imbalanced.After incubating with TSD drug containing serum,endothelial cell proliferation activities and injured cell structures were obviously improved;ET,NO and TGF-beta_1 levels decreased（P0.05,P0.01）,ET/NO ratios approximated to the normal level.Conclusion The main mechanism of TSD for treating ASO ASS lied in improving injured vascular endothelial cells and endocrine disorder.