Watanabe Masanao, Kondo Shoichi, Mizuno Ken, Yano Wataru, Nakao Hiroshi, Hattori Yukio, Kimura Kazuhiro, Nishida Teruo
Tokyo New Drug Research Laboratories, Kowa Company Ltd., Tokyo, Japan.
Invest Ophthalmol Vis Sci. 2006 May;47(5):1862-8. doi: 10.1167/iovs.05-0591.
To investigate the effect of annexin A5, a calcium-dependent phospholipid-binding protein, on corneal epithelial wound healing.
The effect of annexin A5 on migration of rabbit corneal epithelial (RCE) cells in vitro was examined in scrape-wounded cell monolayers. The effect of annexin A5 on the release of urokinase-type plasminogen activator (uPA) from cultured RCE cells was determined by zymography, fluorogenic assay of PA activity, and enzyme-linked immunosorbent assay. The proliferation of RCE cells was assessed by measurement of [3H]thymidine incorporation. The effect of annexin A5 on corneal wound closure in rabbits was investigated after removal of the corneal epithelium, either by exposure to iodine vapor or surgically. Eye drops containing annexin A5 were instilled into one eye and vehicle into the other. The area of the epithelial defect was measured at various times after wounding, and the healing rate was calculated by linear regression analysis.
Annexin A5 significantly promoted the migration of RCE cells in a wounded monolayer. However, annexin A5 had no effect on RCE cell proliferation. Annexin A5 also increased the release of uPA both from wounded RCE cell monolayers and from nonwounded semiconfluent RCE cells. In both models of corneal wound closure, the healing rate was significantly increased by instillation of eye drops containing annexin A5 compared with that apparent in the eyes that received vehicle.
Annexin A5 promoted corneal epithelial wound healing both in vitro and in vivo. Upregulation of uPA release from corneal epithelial cells may contribute to this effect of annexin A5.
研究钙依赖性磷脂结合蛋白膜联蛋白A5对角膜上皮伤口愈合的影响。
在刮伤的细胞单层中检测膜联蛋白A5对兔角膜上皮（RCE）细胞迁移的影响。通过酶谱法、PA活性荧光测定法和酶联免疫吸附测定法测定膜联蛋白A5对培养的RCE细胞中尿激酶型纤溶酶原激活剂（uPA）释放的影响。通过测量[3H]胸苷掺入量评估RCE细胞的增殖。在去除角膜上皮后，通过碘蒸汽暴露或手术的方式，研究膜联蛋白A5对兔角膜伤口闭合的影响。将含有膜联蛋白A5的眼药水滴入一只眼睛，将赋形剂滴入另一只眼睛。在受伤后的不同时间测量上皮缺损面积，并通过线性回归分析计算愈合率。
膜联蛋白A5显著促进受伤单层中RCE细胞的迁移。然而，膜联蛋白A5对RCE细胞增殖没有影响。膜联蛋白A5还增加了受伤的RCE细胞单层和未受伤的半汇合RCE细胞中uPA的释放。在两种角膜伤口闭合模型中，与接受赋形剂的眼睛相比，滴入含有膜联蛋白A5的眼药水显著提高了愈合率。
膜联蛋白A5在体外和体内均促进角膜上皮伤口愈合。角膜上皮细胞uPA释放的上调可能有助于膜联蛋白A5的这种作用。
