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

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)同样的促生长作用。有证据表明，HGF的拮抗剂NK4可以抑制血管内皮生长因子和碱性成纤维细胞生长因子诱导的新生血管形成。(1)检测NK4对人脐静脉内皮细胞和人视网膜色素上皮细胞(ARPE-19)的增殖抑制作用。在96个孔板上，每个孔散布1×10^3个细胞。将其置于浓度为5，10，50,100,500 nm的NK4中72小时。我们用XTT法评价其对细胞增殖的抑制作用。NK4，50 NM以上，与人脐静脉内皮细胞对照。另一方面，任何浓度的NK4都不影响ARPE-19。因此，NK4的细胞增殖抑制作用是对血管内皮细胞的特异性作用，仅抑制新生血管而不影响其周围细胞。(2)评价NK4对实验性脉络膜新生血管的抑制作用。用激光光凝法诱导大鼠实验性新生血管形成，玻璃体腔内注射溶于牛血清白蛋白的5μL液(10μM)。对照组玻璃体腔内注射BSS 5ml。光凝后2周行荧光素血管造影，观察新生血管的形成情况。光镜下测量新生血管厚度。与对照组相比，差异不是决定性的。

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.