人参皂苷Rg3对缺氧诱导的人肾癌786-0细胞VEGF表达的影响及机制探讨

Effect and mechanism of ginsenoside Rg3 on VEGF expression in 786-0 cells induced by hypoxia

目的观察人参皂苷Rg3对缺氧诱导的人肾癌786-0细胞VEGF表达的影响,并探讨其可能的机制。方法将缺氧诱导的786-0细胞分为缺氧组和Rg3组,Rg3组分别加入25、50、100、200μmol/L的Rg3作用24 h,对照组不处理。分别采用RT-PCR法、ELISA法检测786-0细胞中的VEGF mRNA及其蛋白,采用Western blot法检测786-0细胞中的STAT3和MAPKs总蛋白(p38、ERK1/2、JNK)及其蛋白磷酸化水平。结果 Rg3组786-0细胞中VEGF mRNA及其蛋白表达均较对照组显著降低(P均<0.01),且呈浓度依赖性(P均<0.01)。Rg3组786-0细胞中STAT3和MAPKs中ERK1/2、JNK蛋白磷酸化水平较对照组显著降低(P均<0.01),且呈浓度依赖性(P均<0.01),STAT3和p38、ERK1/2、JNK、pp38水平无明显差异。结论缺氧状态下Rg3可抑制786-0细胞的VEGF表达,其机制与抑制STAT3和MAPKs蛋白磷酸化有关。

Objective To investigate the effect and mechanism of ginsenoside Rg3 on VECF expression in 786-0 cells induced by hypoxia.Methods 786-0 cells were cultured with Rg3 in normoxia and hypoxia conditions,and were divided into Rg3 group and control group.786-0 cells in Rg3 group were cultured with 25、50、100、200μmoL/L Rg3 for 24 h.Real -time PCR was used to detect VEGF mRNA expression,ELISA was used to detect VEGF protein secretion,Western blot was used to detect STAT3,MAPKs(p38,ERK1/2,JNK) and protein phosphorylation.Results Compare with control group,VEGF mRNA and protein expression,STAT3 and MAPKs(ERK1/2,JNK) protein phosphorylation in Rg3 group786- 0 cells decreased in a concentration-dependent manner.Conclusion Rg3 can inhibite VECF expression in normoxia and hypoxia conditions,it may be related with the inhibition of STAT3 and MAPKs protein phosphorylation.