Wang Mei-jian, Du Dan-yu, Fan Wei, Zhang Cang, Liu Yang, Fan Jia-hong, Yuan Sheng-tao, Lin Sen-sen
Yao Xue Xue Bao. 2016 Feb;51(2):309-15.
This study was designed to investigate the effect of Xiao-Ai-Ping injection on cancer angiogenesis. CCK8 assay and Brd U incorporation immunofluorescence assay were used to detect the effect of Xiao-Ai-Ping injection on HUVECs proliferation; wound healing assay and transwell assay were employed to test the effect of Xiao-Ai-Ping injection on HUVECs migration. The anti-angiogenic effect of Xiao-Ai-Ping injection was examined by tube formation assay, rat aortic ring assay and chicken chorioallantoic membrane(CAM) assay. ELISA assay was used to measure the secretion of vascular endothelial growth factor(VEGF); and the activation of vascular endothelial growth factor receptor 2(VEGFR2) protein and its downstream signaling pathways were examined by Western blot. Our data demonstrated that Xiao-Ai-Ping injection inhibited HUVECs proliferation in a time- and dose-dependent manner, and the IC(50) (mg·m L(-1)) values for 24, 48 and 72 h were 48.7 ± 7.14, 29.1 ±2.25 and 22.0 ± 4.53, individually. Xiao-Ai-Ping injection inhibited HUVECs DNA synthesis and migration. Xiao-Ai-Ping injection suppressed HUVECs tube formation, and reduced microvessel sprouting from rat aortic rings and vessel growth in CAMs. Furthermore, Xiao-Ai-Ping injection attenuated the secretion of VEGF, and inhibited the expression of p-VEGFR2 and phosphorylation of protein kinase B(p-AKT). We conclude that Xiao-Ai-Ping injection inhibits angiogenesis by down-regulation of VEGF signaling and AKT pathway.
本研究旨在探讨消癌平注射液对肿瘤血管生成的影响。采用CCK8法和BrdU掺入免疫荧光法检测消癌平注射液对人脐静脉内皮细胞（HUVECs）增殖的影响；采用划痕愈合实验和Transwell实验检测消癌平注射液对HUVECs迁移的影响。通过管腔形成实验、大鼠主动脉环实验和鸡胚绒毛尿囊膜（CAM）实验检测消癌平注射液的抗血管生成作用。采用ELISA法检测血管内皮生长因子（VEGF）的分泌；通过蛋白质免疫印迹法检测血管内皮生长因子受体2（VEGFR2）蛋白的激活及其下游信号通路。我们的数据表明，消癌平注射液以时间和剂量依赖性方式抑制HUVECs增殖，24、48和72小时的半数抑制浓度（IC50）（mg·mL-1）值分别为48.7±7.14、29.1±2.25和22.0±4.53。消癌平注射液抑制HUVECs DNA合成和迁移。消癌平注射液抑制HUVECs管腔形成，并减少大鼠主动脉环微血管的出芽和CAMs中的血管生长。此外，消癌平注射液减弱了VEGF的分泌，并抑制了p-VEGFR2的表达和蛋白激酶B（p-AKT）的磷酸化。我们得出结论，消癌平注射液通过下调VEGF信号通路和AKT途径抑制血管生成。
