加减大黄虫丸抑制血管生成的实验研究

Experimental Study on Modified "Dahuang Zhechong Pill" in Inhibiting Angiogenesis

目的探讨加减大黄虫丸对血管生成的抑制作用。方法鸡胚绒毛尿囊膜(CAM)法检测加减大黄虫丸对血管生成的影响;采用MTS比色法观察不同浓度的加减大黄虫丸对血管内皮生长因子(VEGF)诱导的ECV-304细胞增殖的影响;Tran-swell小室检测不同浓度的加减大黄虫丸对ECV-304细胞移行的影响;Matrigel实验检测不同浓度的加减大黄虫丸对ECV-304细胞内皮管腔形成的影响。结果加减大黄虫丸中、低浓度组能够抑制鸡胚CAM血管生成;MTS比色法显示,0.05-0.20g/m 浓度的加减大黄虫丸对VEGF诱导的ECV-304细胞增殖具有抑制作用,而更低和更高浓度的加减大黄虫丸则无抑制作用。Transwell小室实验显示,加减大黄虫丸浓度为0、12.5、25.0和50.0mg/m l时ECV-304细胞移行数分别为208.67±17.16、132.67±16.50、78.33±13.50和18.67±6.66;M atrigel实验显示,加减大黄虫丸浓度为0、12.5、25.0和50.0mg/m l时ECV-304细胞内皮管腔形成数分别为25.67±1.53、22.33±1.53、16.33±2.52和2.33±1.53,可见抑制细胞移行和管腔形成的作用随浓度增大而增强。结论加减大黄虫丸具有明显抑制血管生成的作用。

Objective To investigate the effect of modified ＂ Dahuang Zhechong Pill＂ on angiogenesis. Methods Chick embryo chorioallantoic membrane （CAM） method was used to assess the effect of modified ＂Dabuang Zhechong Pill＂ on angiogenesis. MTS method was used to assess the effect of modified ＂Dahuang Zhechong Pill＂ at different concentrations on the proliferation of ECV-304 cells induced by vascular endothelial growth factor（VEGF）. Transwell insert was used to as- sess the effect of modified ＂ Dahuang Zhechong Pill＂ at different concentrations on the migration of ECV-304 cells. Matrigel was used to assess the effect of modified ＂Dahuang Zhechong Pill＂ at different concentrations on the tube formation of ECV-304 cells. Results modified ＂Dahuang Zhecbong Pill＂ at median and low concentrations could inhibit the angiogenesis of chick embryo CAM. The proliferation of ECV-304 cells induced by VEGF was inhibited by modified ＂Dahuang Zhechong Pill＂ at concentrations from 0.05 g/mt to 0.2 g/ml, while it wasn＇ t inhibited at higher or lower concentrations. When the concentrations of modified ＂Dahuang Zhechong Pill＂ were 0, 12.5,25, and 50 mg/ml, cell numbers that migrated to under the Transwell membrane were respectively 208.67 ± 17.16, 132.67 ± 16, 50,78.33 ± 13.50, and 18.67 ± 6.66. When the concentrations of modified ＂Dahuang Zhechong Pill＂ were 0, 12.5, 25, and 50 mg/ml, tube numbers formationed in Matrigel were respectively 25.67 ± 1.53, 22.33 ± 1.53, 16.33 ±2.52 and 2.33 ± 1.53. Conclusion Modified ＂Dahuang Zhechong Pill＂ is notably effective in inhibiting angiogenesis.