扶正解毒方选择性抑制肿瘤血管生成及调控VEGF/VEGFR-2信号通路的实验研究

Studies on Selective Inhibition of Fuzheng Jiedu Fang (扶正解毒方) on Tumor Angiogenesis and Regulation on VEGF/VEGFR-2 Signal Transduction Pathway

目的:探讨扶正解毒方选择性抑制肿瘤血管生成及其相关作用机制。方法:以肿瘤血管内皮细胞模型、正常血管内皮细胞及Lewis肺癌荷瘤小鼠为研究对象,随机分为生理盐水组、贝伐单抗组、扶正解毒组及其拆方扶正、解毒药物组,采用MTT法观察扶正解毒方药、贝伐单抗对肿瘤血管内皮细胞与正常血管内皮细胞增殖的影响;采用免疫组化法观察药物对Lewis肺癌荷瘤小鼠瘤组织VEGF表达的影响;采用Western Blotting法观察药物对血管内皮细胞VEGFR-2(KDR)表达的影响。结果:扶正解毒方能够抑制肿瘤血管内皮细胞增殖(P<0.05),而对正常血管内皮细胞增殖没有明显影响(P>0.05),贝伐单抗对肿瘤血管内皮细胞及正常血管内皮细胞增殖均有抑制作用(P<0.01),扶正解毒组与贝伐单抗组比较有明显差异(P<0.05);扶正解毒方能降低瘤组织VEGF表达及肿瘤血管内皮细胞VEGFR-2表达(P>0.05),扶正解毒方组VEGFR-2表达水平与正常血管内皮细胞相近。结论:扶正解毒方能够选择性抑制肿瘤血管生成,其作用机制与调控VEGF/VEGFR-2信号传导有关。

Objective： To investigate the selective inhibiton of Fuzheng Jiedu Fang（ FZJDF） on tumor angiogenesis and regulation in signal transduction. Methods： Tumor- derived endothelial cells（ Td- ECs） and human umbilical vein endothelial cells（ HUVECs） were cultured in vitro. Td- ECs,HUVECs and mice were randomly divided into 5 groups： saline group,FZ group,JD group,FZJDF group and Bevacizumab group. Treatment was as scheduled and the tumors were carefully removed. The inhibitory effect of FZJDF and Bevacizumab on Td- ECs and HUVECs was tested by MTT method. The expression of VEGF in implanted tumor of mice was tested by immunohistochemistry. The expression VEGFR- 2 of Td- ECs was tested by Western blotting method. Results：FZJDF could inhibit the proliferation of Td- ECs（ P〈0. 05）,but those showed no direct inhibition on the proliferation of HUVECs（ P〈0. 05）. Bevacizumab group could obviously inhibit the proliferation of Td- ECs and HUVECs（ P〈0. 01）. Compared with the FZJD group,the difference was significant（ P〈0. 05）. FZJDF could lower the expression of VEGF in tumor and VEGFR- 2 of Td- ECs. The expression levels of VEGFR- 2changed little between FZJD group and HUVECs group（ P〈0. 05）. Conclusion： FZJDF could selectively inhibit tumor angiogenesis,the mechanism of which is the regulation of VEGF / VEGFR- 2 pathway.