三棱、莪术含药血清对培养的人脐静脉血管内皮细胞生长和VEGF表达的影响

Effect of medicated serum of Sangleng and Eshu on human umbilical vein vascular endothelial growth factor and vascular endothelial cells

目的探讨三棱、莪术含药血清对血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)诱导的血管内皮细胞增殖和VEGF表达影响。方法以三棱、莪术含药血清作用于VEGF诱导的人脐静脉血管内皮细胞(humanumbili-calveinendothelialcell-1,HUVEC-1),倒置相差显微镜观察细胞形态学改变,MTT法观察细胞增殖情况,Westernblot和RT-PCR方法分别检测内皮细胞VEGF蛋白和mRNA的表达。结果三棱、莪术5.0g·kg-1·d-1、2·5g·kg-1·d-1大鼠含药血清可使正常人脐静脉血管内皮细胞排列紊乱,明显梭形化。5.0g·kg-1·d-1大鼠10%、5%、2.5%含药血清组和2.5g·kg-1·d-1大鼠10%含药血清组HUVEC-1细胞的增殖显著低于空白血清相同浓度组(P<0·05)。5.0g·kg-1·d-1、2.5g·kg-1·d-1大鼠5%的含药血清使HUVEC-1细胞VEGF蛋白、VEGFmRNA表达均显著降低。结论三棱、莪术含药血清可抑制VEGF诱导的血管内皮细胞增殖,其机制与抑制血管内皮生长因子表达密切相关。

Objective To explore the effect of medicated serum of Sangleng and Eshu on the expression of vascular endothelial growth factor and vascular endothelial cell proliferation induced by VEGF in vitro. Methods Medicated serum of Sangleng and Eshu was used to culture human umbilical vein endothelial cell （HUVEC-1 ） induced by VEGF. The morphologic changes of HUVEC-1 were observed with phase contrast microscope, and cell proliferation was detected by MTT method, and the expression of vascular endothelial growth factor protein and mRNA in endothelial cells was detected by Western blotting and RT-PCR. Results The medicated serum of 5.0, 2.5 g·kg^-1·d^-1 Sangleng and Eshu could cause arrangement disorder in the normal umbilical vein endothelial cells. The medicated serum of 5. 0 g·kg^-1·d^-1 Sangleng and Eshu （10%, 5%, 2.5% ） and medicated serum of 2.5 g·kg^-1·d^-1 （10%） could inhibit vascular endothelial cell proliferation remarkably （ P 〈 0.05 ）, as compared with the blank serum of identical concentration. The medicated serum of 5 g ·kg^-1·d^-1 and 2.5 g·kg^-1·d^-1 （5%） could reduce significantly the expression of vascular endothelial growth factor protein and mRNA of HUVEC-1. Conclusion The medicated serum of Sangleng and Eshu could inhibit vascular endothelial cell proliferation induced by VEGF in vitro, and its mechanism might be associated with inhibited expression of vascular endothelial growth factor.