血小板反应素-1对猪胸导管内皮细胞增殖和凋亡的影响

The Effect of Thrombospondin-1 on the Proliferation and Apoptosis of Lymphatic Endothelial Cells in Pig

目的探讨血小板反应素-1(thrombospondin-1,TSP-1)对淋巴管内皮细胞(lymphatic endothelial cells,LECs)增殖和凋亡的影响。方法取健康猪的胸导管内皮细胞进行分离、培养;应用凝血因子VIII,血管内皮生长因子受体-3(VEGFR-3)抗体联合对淋巴管内皮细胞进行鉴定;采用MTT法检测TSP-1对LEC的生长抑制作用;采用Hoechst33258荧光染色检测淋巴管内皮细胞凋亡。结果培养的淋巴管内皮细胞呈第VIII因子和VEGFR-3阳性反应,为典型淋巴管内皮细胞。MTT法证实,当浓度为0.8μg/ml￣2.0μg/ml时,TSP-1能明显抑制淋巴管内皮细胞的增殖,且抑制作用呈现剂量依赖关系。Hoechst33258证实,经TSP-1处理后淋巴管内皮细胞,可观察到其核周围有凋亡小体。结论TSP-1对淋巴管内皮细胞增殖有明显的剂量依赖性抑制作用,诱导淋巴管内皮细胞凋亡。

Objective To explore the effect of thrombospondin-1 （TSP-1） on lymphatic endothelial cells （LECs） proliferation and apoptosis in pig. Methods LECs ducted from healthy pigs were cultured, antibodies against blood coagulation factor VIII and vascular endothelial growth factor receptor-3（VEGFR-3） were used to identify LCEs. The MTT was used to examine the effectiveness of TSP-1 on LECs proliferation, Hoechst33258 fluorescence staining was used to detect the apoptotic endothelial cells. Results Cultured LECs were both blood coagulation factor VIII positive and VEGFR-3 positive. MTT results indicated that 0.8 μg/ml -2.01xg/ml TSP-1 could obviously inhibit the proliferation of LECs with dose-dependent matter, Hoechst33258 staining confirmed that apoptotic bodies could be observed in LECs treated by TSP-1. Conclusion TSP-1 could evidently inhibit the proliferation of LECs dose-dependently and induce the apoptosis of LECs.