人参皂甙Rg3对淋巴管内皮细胞生成的影响

Influence of 20(R)-ginsenoside Rg3 on lymphangenesis

目的探讨人参皂甙Rg3[20(R)-Ginsenoside Rg3]对淋巴管内皮细胞迁移、增殖能力及凋亡的影响。方法用含不同浓度人参皂苷Rg3的培养液培养Hela细胞,收集培养上清并制备条件培养液(CM);取健康猪的胸导管内皮细胞进行分离、培养;Ⅷ因子、VEGFR-3抗体联合对淋巴管内皮细胞进行鉴定;通过划线刮除法和MTT法观察人参皂甙Rg3对淋巴管内皮细胞的迁移和增殖能力的影响;Hoechst33258荧光染色检测淋巴管内皮细胞凋亡。结果第Ⅷ因子和VEGFR-3抗体对培养的淋巴管细胞进行联合鉴定,为典型淋巴管内皮细胞;应用划线刮除法和MTT法对对照组与实验组进行比较显示:不同浓度的人参皂甙Rg3能够明显抑制淋巴管内皮细胞的增殖和游走,差异有统计学意义(P<0.01);Hoechst33258证实,经人参皂甙Rg3条件培养液处理后淋巴管内皮细胞,可观察到其核周围有凋亡小体。结论人参皂甙Rg3对淋巴管内皮细胞的迁移和增殖能力有明显的抑制作用,并且有剂量依赖性;人参皂甙Rg3能够诱导淋巴管细胞凋亡。

[Abstract] Objective To explore the effect of 20 （R）-Ginsenoside Rg3 on lymphatic endothelial cell migration, proliferation and apoptosis. Methods Hela cells were cultured in media containing Rg3, and conditioned media （CM） were prepared with the culture media from hela cells; LECs which ducted from healthy pig were cultured; Ⅷ and VEGFR-3 were used to idenifty LCEs. The methods of craped line and MTT were used to examine the effectiveness of 20（R）-Ginsenoside Rg3 on lymphatic endothelial cell migration and proliferation; Hoechst33258 fluorescence staining was used to detect endothelial cell apoptosis. Results Ⅷ factor and VEGFR-3 antibody were used to identify the cultured lymphatic cells. The result showed that they were typical lymphatic endothelial cells; The scraped line and MTT method were used to compare the difference between the control group and the experimental group. The result showed： different concentrations of 20（R）-Ginsenoside Rg3 can inhibit lymphatic endothelial cell proliferation and migration, the difference was significant （P〈0.01）; Hoechst33258 staining confirmed that apoptotic bodies could be observed around its nuclear in 20 （R）-Ginsenoside Rg3 treated LECs. Conclusion 20（R）- Ginsenoside Rg3 could evidently inhibit the migration and proliferation of lymphatic endothelial cells and the effects were dosedependent; 20（R）-Ginsenoside Rg3 could induce the apoptosis of LECs.