人参皂苷Rg3抑制急性白血病骨髓基质细胞HIF-1α及VEGF的表达及其机制探讨

Ginsenosisde Rg3 inhibits HIF-1α and VEGF expressions in acute leukemia bone marrow stromal cells

目的探讨人参皂苷Rg3对急性白血病骨髓基质细胞HIF-1α及VEGF表达的作用及其可能的机制。方法培养正常人及急性白血病患者的骨髓基质细胞,MTT法检测人参皂苷Rg3对正常及急性白血病骨髓基质细胞增殖的抑制作用;RT-PCR检测人参皂苷Rg3处理前后急性白血病骨髓基质细胞HIF-1α及VEGF mRNA表达的变化;Western blot、免疫荧光检测人参皂苷Rg3处理前后急性白血病骨髓基质细胞HIF-1α、VEGF、p-Akt、p-ERK蛋白表达的变化。结果人参皂苷Rg3对骨髓基质细胞增殖抑制的最适作用时间为24h,最适作用浓度为40μg/ml;处理24h后,急性白血病骨髓基质细胞HIF-1α、VEGF mRNA表达及蛋白表达均明显减少(P<0.05);p-Akt及p-ERK蛋白表达亦较处理前明显减少(P<0.05)。结论人参皂苷Rg3可明显抑制急性白血病骨髓基质细胞HIF-1α、VEGF mRNA及蛋白水平的表达,同时可抑制与血管形成密切相关的MAPK、PI3K/Akt途径中Akt、ERK蛋白的磷酸化,提示人参皂苷Rg3可能通过阻断PI3K/Akt、MAPK信号途径抑制急性白血病骨髓的血管生成。

Objective To investigate the effect of ginsenoside Rg3 on HIF-1α and VEGF expressions in acute leukemia bone marrow stromal cells（BMSCs）and the possible underlying mechanism.MethodsBMSCs from normal health individuals and acute leukemia patients were cultivated.The appropriate time and concentration of ginsenoside Rg3 on BMSCs＇ proliferation were assessed by MTT.The BMSCs were treated with ginsenoside Rg3,and mRNA levels of HIF-1α and VEGF were detected by RT-PCR.In addition,the expre-ssion of HIF-1α,VEGF,p-Akt and p-ERK proteins were measured by Western blot analysis and immunofluorescence assays.ResultsAfter the addition of ginsenoside Rg3,MTT showed that the appropriate concentration and time was 40 μg/ml and 24 h respectively.HIF-1α and VEGF expressions at mRNA and protein levels were reduced significantly（P〈0.05）.The protein expressions of p-Akt and p-ERK were decreased significantly（P〈0.05）.ConclusionGinsenoside Rg3 not only significantly inhibits HIF-1α and VEGF mRNA expression but also protein expression in acute leukemia BMSCs.It also down-regulates the phosphorylation of Akt and ERK in PI3K/Akt and MAPK pathways respectively,which are involved in angiogenesis.These results demonstrate an anti-angiogenesis function of ginsenoside Rg3 through PI3K/Akt and MAPK signal pathways in acute leukemia bone marrow.