蛇毒金属蛋白酶抑制剂重组蛋白抗肿瘤血管生成作用及其机制

发布时间：2018-03-26 00:03

本文选题：蛇毒　切入点：基质金属蛋白酶抑制剂　出处：《中国药理学通报》2017年03期

【摘要】：目的研究蛇毒金属蛋白酶抑制剂重组蛋白(recombinant snake venom metalloproteinase inhibitor,r SVMPI)对血管生成的影响及其分子机制。方法应用鸡胚绒毛尿囊膜(chicken chorioallantoic membrane,CAM)血管生成模型观察SVMPI重组蛋白对血管生成的影响;采用Alamar Blue分析方法检测人脐静脉内皮细胞(human umbilical veins endothelial cells,HUVECs)增殖能力、Annexin V-FITC双标记流式细胞术检测细胞凋亡、划痕标记法检测细胞体外迁移能力、Boyden小室分析方法检测细胞体外趋化能力及管腔形成法检测体外血管新生能力;通过real-time PCR及Western blot检测重组蛋白处理后的HUVECs KDR、FGFR-1表达。结果r SVMPI减少鸡胚尿囊膜新生血管密度指数,减弱由VEGF诱导的HUVECs趋化能力,抑制HUVECs体外新生小管的形成,降低HUVECs细胞KDR和FGFR-1的表达水平。结论r SVMPI可能通过阻断VEGF-KDR或b FGF-FGFR信号转导途径,发挥抑制血管生成的作用。
[Abstract]:Objective to study the effect of recombinant snake venom metalloproteinase inhibitor (SVMPI) on angiogenesis and its molecular mechanism. Methods the angiogenesis model of chicken chorioallantoic membrane was used to observe the effect of SVMPI recombinant protein on angiogenesis. Alamar Blue assay was used to detect the proliferation of human umbilical veins endothelial cells (HUVECs) and Annexin V-FITC double labeling flow cytometry (FCM) was used to detect the apoptosis of human umbilical vein endothelial cells (HUVECs). In vitro migration ability was detected by scratch labeling method. Boyden chamber analysis method was used to detect the chemotaxis of cells in vitro and angiogenesis in vitro by lumen formation method. The expression of HUVECs KDRN FGFR-1 was detected by real-time PCR and Western blot. Results r SVMPI reduced the angiogenesis density index of chicken embryo allantoic membrane, decreased HUVECs chemotaxis induced by VEGF, and inhibited the formation of new tubules of HUVECs in vitro. Conclusion r SVMPI may inhibit angiogenesis by blocking VEGF-KDR or b FGF-FGFR signal transduction pathway.
【作者单位】：福建医科大学附属第一医院皮肤科;福建医科大学分子医学研究中心消化道恶性肿瘤教育部重点实验室;福建省肿瘤医院病理科;
【基金】：福建省卫生系统中青年骨干人才培养项目(No 2014-ZQNJC-17)
【分类号】：R96

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