土贝母苷甲对人脐静脉内皮细胞凋亡和肿瘤诱导的血管生成的影响

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研究了土贝母苷甲(下称苷甲)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)凋亡和肿瘤诱导的血管生成的影响。包括:MTT法检测苷甲对HUVECs生长的影响;荧光显微镜观察苷甲作用下HUVECs的形态变化;流式细胞术分析苷甲对HUVECs周期及凋亡的影响;聚碳酸酯膜小室(Transwell model)趋化运动模型检测苷甲对HUVECs运动能力的影响;鸡胚绒毛尿囊膜(chick embryochorioallantoic membrane,CAM)试验检测苷甲对人鼻咽癌细胞(human nasopharyngeal carcinomaCNE-2Zcells,CNE-2Z)诱导的CAM血管生成的影响;免疫组化法检测苷甲对BALB/c裸小鼠Lewis肺癌组织微血管密度(microvessel density,MVD)和血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)和血小板源生长因子(platelet-derived growth factor,PDGF)表达的影响。苷甲明显抑制HUVECs的生长,其抑制效果与剂量和作用时间相关,苷甲作用HUVECs24、48、72h其IC50值分别为24.2、21.4、17.9μmol/L;苷甲作用下HUVECs发生周期阻滞,呈现典型的凋亡特征,20.0μmol/L苷甲作用12、24、36hHUVECs的凋亡率分别为11.4%、20.8%、25.3%;20.0μmol/L苷甲处理HUVECs24h,对细胞迁移抑制率为58.4%;苷甲抑制CNE-2Z细胞诱导的CAM血管生成,并与剂量相关;苷甲应用后瘤组织MVD明显减少,VEGF、bFGF、PDGF表达下调。苷甲有明显的抑制血管生成活性,其抑制血管生成作用与诱导血管内皮细胞凋亡,抑制其运动能力,下调VEGF、bFGF和PDGF的表达有关。 The effects of saponin A on the apoptosis and tumor-induced angiogenesis of human umbilical endothelial cells (HUVECs) were studied. Including: The effect of glycosides on the growth of HUVECs was detected by MTT assay; The morphological changes of HUVECs were observed by fluorescence microscopy; The effects of glycosides on the cycle and apoptosis of HUVECs were analyzed by flow cytometry; Transwell model A chemotaxis model was used to detect the effect of glycosides on the exercise performance of HUVECs. The chicken embryo chorioallantoic membrane (CAM) assay was used to detect glycosides induced by human nasopharyngeal carcinoma CNE-2Z cells (CNE-2Z). Effects of CAM on angiogenesis; Immunohistochemical method for the detection of glycoside A in BALB/c nude mice Lewis lung cancer Tissue microvessel density (MVD) and vascular endothelial growth factor (VEGF), alkaline fibroblasts Effects of basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) expression. The glycoside significantly inhibited the growth of HUVECs. The inhibitory effect was related to dose and duration of action. The IC50 values ​​of 24, 48, and 72 h after administration of glycosides were 24.2, 21.4, and 17.9 μmol/L, respectively; HUVECs were blocked by glycoside A. The typical apoptotic characteristics were observed. The apoptotic rates of HUVECs were 12.4%, 20.8%, and 25.3% at 2, 24, and 36 h after treatment with 20.0 μmol/L glycosides, respectively. The HUVECs treated with 20.0 μmol/L glycosides for 24 h had 58.4 inhibition of cell migration. %; glycosides inhibited the angiogenesis of CAM induced by CNE-2Z cells, and correlated with the dose; MVD of tumor tissue was significantly decreased and the expression of VEGF, bFGF and PDGF was down-regulated after application of glycosides. The glycoside has obvious inhibitory activity on angiogenesis. It inhibits angiogenesis and induces apoptosis of vascular endothelial cells, inhibits their ability to exercise, and down regulates the expression of VEGF, bFGF and PDGF.
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