雷公藤内酯醇对外周血内皮祖细胞的影响

Effects of triptolide on changes of endothelial progenitor cells from peripheral blood

目的:观察雷公藤内酯醇对人外周血内皮祖细胞(endothelial progenitor cells,EPCs)的影响。方法:密度梯度离心法获取人外周血单个核细胞,接种至人纤维连接蛋白包被的培养板,培养7 d后收集贴壁细胞进行细胞免疫化学鉴定。实验加入不同浓度雷公藤内酯醇(2.5μg/L,5.0μg/L,10.0μg/L,20.0μg/L)干预24 h,采用MTT比色法、改良的Boyden小室和黏附能力测定实验,分别观察EPCs的增殖、迁移和黏附能力。结果:雷公藤内酯醇显著影响EPCs的各项生物学功能,使EPCs的黏附能力显著减弱,迁移功能显著下降,增殖能力显著被抑制,这些改变均存在明显的浓度依赖性,20μg/L浓度组达最大抑制;在雷公藤内酯醇低浓度组,冠心病患者EPCs细胞的各项功能明显低于非冠心病患者组,高浓度组两组相近。结论:雷公藤内酯醇可能通过抑制EPCs增殖、迁移和黏附功能,抑制血管再内皮化和血管新生。

AIM： To investigate the effects of triptolide on endothelial progenitor ceils from peripheral blood. METHODS ： Total mononuclear cells （MNCs） were isolated from peripheral blood by density gradient centrifugation. The cells were plated on fibronectin - coated culture dishes. After 7 d of culture, adherent cells were characterized by demonstrating the expression of CD34, CD31 and vWF with immunohistochemistry. Adherent cells were stimulated with triptolide （2. 5, 5.0, 10. 0, 20. 0 txg/L） or vehicle control for 24 h. Activities of EPCs in terms of proliferation and migration were determined by MTF assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating MNCs on fibronectin -coated dishes, and then the adherent cells were counted. RESULTS： The proliferative, migratory and adhesive capacities of EPCs decreased significantly after 24 h incubation with triptolide, maximum at 20 μg/L （ compared to that in control group）. In patients with coronary heart disease, the biological function of EPCs was lower than that in patients without coronary heart disease in low dosage of triptolide but almost the same in high dosage group. CONCLUSION： Triptolide may inhibit functional activities of EPCs, the reendothelization＇and neovascularization of vessel.