当归补血汤抑制与肿瘤共培养血管内皮细胞的增殖及其分子机制

目的:通过当归补血汤对与肿瘤共培养血管内皮细胞增殖及分子表达的研究,探讨当归补血汤抑制肿瘤血管生成的可能机制。方法:应用Transwell建立与肿瘤共培养血管内皮细胞模型(以下简称共培养模型),在不同剂量当归补血汤的干预下,通过CCK-8法观察与肿瘤共培养血管内皮细胞的增殖作用;通过ELISA法检测与肿瘤共培养血管内皮细胞血管内皮生长因子(VEGF)及其受体(VEGFR1,VEGFR2,sVEGFR1,sVEGFR2)的表达。结果:当归补血汤能够促进正常血管内皮细胞增殖,与正常组比较,高、中、低剂量组均有差异性(P<0.01,P<0.01,P<0.05);抑制与肿瘤共培养血管内皮细胞增殖,且与剂量呈正相关,与共培养模型组比较,各剂量组均有显著差异(P<0.01)。当归补血汤各剂量组均能促进与肿瘤共培养血管内皮细胞VEGF的表达;抑制与肿瘤共培养血管内皮细胞VEGFR1,VEGFR2的表达,且与剂量呈正相关;促进与肿瘤共培养血管内皮细胞sVEGFR1,sVEGFR2的表达。结论:当归补血汤能够抑制与肿瘤共培养血管内皮细胞的增殖,其机制可能与其调节VEGF与VEGFR和sVEGFR两种受体的结合有关。

Effect of Danggui Buxue Decoction on Proliferation and its Molecular Mechanism in Tumor Vascular Endothelial Cells

Objective: To study the effect of Danggui Buxue decoction(DBD) on proliferation in tumor vascular endothelial cells,and to explore the mechanism of inhibition of tumor angiogenesis. Method: After different doses of DBD acting on vascular endothelial cells (EA. Hy926) co-cultured with the tumor cells (A549) model, CCK-8 assay was used to observe the proliferation of vascular endothelial cells co-cultured with the tumor cells; enzyme-cinked immunosorbant assay (ELISA) was used for assay of vascular endothelia growth factor (VEGF) and its receptors (VEGFR1, VEGFR2, sVEGFR1, sVEGFR2) and expression of vascular endothelial cells co-cultured with the tumor cells. Result: Compared with normal group, vascular endothelial cells co-cultured with tumor cells proliferation capacity increased (P<0.01); DBD each dose group could promote the proliferation of normal endothelial cells, inhibit the proliferation of vascular endothelial cells co-cultured with tumor cells with a manner of dose and effect, compared with model group, there was significant difference (P<0.01). DBD each dose could promote vascular endothelial cell co-cultured with tumor cells expression of VEGF, compared with model group, co-cultured of high dose group and dose group showed a significantly different (P<0.01). Co-culture model group was significantly up-regulate VEGFR1,VEGFR2 expression, the groups DBD inhibited the expression of VEGFR1,VEGFR2 and showed a dose relationship. Co-cultured with tumor vascular endothelial cells in model group decreased expression of sVEGFR1 and sVEGFR2, each dose group of DBD could promote sVEGFR1 and sVEGFR2 expression,showed a dose relationship. Conclusion: DBD can Inhibit proliferation of vascular endothelial cells co-cultured with the tumor cells. The mechanism might be related to regulation of VEGF and its receptors