The trial of treatment in choroidal neovascularization by the drug release system of NK4,inhibitors of hepatocyte growth factor

NK4、肝细胞生长因子抑制剂药物释放系统治疗脉络膜新生血管的试验

• 批准号: 14571682
• 负责人: OKUDA Masatoshi
• 金额: $ 2.24万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571682/
• 关键词: NK4; HGF; Biodegradable scleral implant; Drug delivery system; HUVEC; CNV; 生体分解性高分子; 生体非分解性高分子; リン酸ベタメサゾン; ベタメサゾン; 強膜; 肝細胞増殖因子阻害剤; 脈絡膜新生血管; ARPE-19

Hepatocyte growth factor (HGE) is shown growth promoting action as much as VEGF (vascular endothelial growth factor) or bFGF (basic fibroblast growth factor) to the human umbilical vein endothelial cells (HUVEC). It is becoming clear that NK4,which is the antagonist of the HGF, inhibits the neovascularization was induced VEGF and bFGF.(1)We evaluated the proliferating suppression effect of NK4 to the HUVEC and the human retinal pigment epithelial cell (ARPE-19). On the 96 well plates, each hole was scattered 1x10^3 each cells. It was made to expose for 72 hours to the NK4 of the concentration of 5,10,50,100,500 nM. We evaluated the cellular proliferating suppression effect by using a XTT assay. NK4,50 nM or more, contrastively controlled to HUVEC. On the other hand, Any concentration of NK4 was not influenced ARPE-19. Therefore, it was suggested that a cellular proliferation suppression effect of NK4 was specific action to vascular endothelial cells and controlled only neovascularization without affecting its surrounding cells.(2)We evaluated the suppression effect of NK4 to the experimental choroidal neovascularization. Experimental CNV was induced in rats by laser photocoagulation, and 5μl (10μM) of NK4,dissolved in BSS, was injected in the vitreous cavity. Controls received a intravitreal injection 5ml of only BSS. We performed fluorescein angiography 2nd weeks after photocoagulation, and we evaluated the formation of CNV. It measured the thickness of the CNV with light microscope. As compared with contrast, the difference was not seen conclusively.

肝细胞生长因子（HGE）对人脐静脉内皮细胞（HUVEC）具有与血管内皮生长因子（VEGF）或碱性成纤维细胞生长因子（bFGF）同等的促生长作用。越来越清楚的是，NK4作为HGF的拮抗剂，抑制了VEGF和bFGF诱导的新生血管形成。(1)评价NK4对HUVEC和人视网膜色素上皮细胞（ARPE-19）的增殖抑制作用。在96孔板上，每个孔每个细胞分散1 × 10^3。分别在5、10、50、100、500 nM浓度的NK4中暴露72小时。我们用XTT法评估细胞增殖抑制作用。NK4,50 nM以上，与HUVEC对比控制。另一方面，NK4的浓度对ARPE-19没有影响。因此，我们认为NK4的细胞增殖抑制作用是对血管内皮细胞的特异性作用，仅控制新生血管而不影响其周围细胞。(2)我们评价了NK4对实验性脉络膜新生血管的抑制作用。采用激光光凝法诱导大鼠实验性CNV，将溶解于BSS中的NK4 5μl （10μM）注射于玻璃体腔内。对照组只接受5ml的BSS玻璃体内注射。光凝后第2周行荧光素血管造影，观察CNV的形成情况。用光镜测量CNV的厚度。与对照相比，这种差异并不能确定。