作者：王冰 作者单位：山东大学 附属省立医院眼科,济南 250021
【摘要】 目的 研究细胞外基质受体Ⅲ(CD44)及其配体透明质酸(HA)在角膜碱烧伤修复过程中的作用。 方法 制作角膜碱烧伤大鼠模型,免疫组织化学方法观察CD44及HA在角膜碱烧伤后的表达变化,以正常大鼠和机械性划伤大鼠角膜为对照。 结果 CD44及HA在正常及机械性伤大鼠角膜仅少量表达；碱烧伤模型中烧伤后12 h即可见表达增多,同时角膜鳞状上皮细胞、翼状细胞出现CD44。 结论 在角膜碱烧伤过程中CD44及HA对角膜的重新上皮化可能有积极作用。
【关键词】 抗原 CD44 角膜
Expression of CD44 and Hyaluronan in the Alkalineburned Corneal of Rat
WANG Bing, ZHU Xuejun, XU Guoxing, WANG Lihua
1.Department of Ophthalmology, The Affiliated Shandong Provincial Hospital,
Shandong University,Jinan 250021,China;
2.Department of Ophthalmology,The Affiliated First Hospital,
Fujian Medical University,Fuzhou 350005,China
ABSTRACT: Objective To investigate the expression of CD44 and hyaluronan in the corneal of alkalineburnedrat (ABR) model and to reveal the function of them in the case of corneal rehabilitation. Methods The ABR model of the corneal was first developed, monitor the expression of CD44 and hyaluronan at the time point of 12 h,24 h,3 d,7 d,14 d after the injury was monitored by the means of immunohistochemical analysis. Results In the corneas of control teams, CD44 was little only expressed only in the basal column cells and the stromal kerotocytes at a low level. At the time point of 12 h after injury, increaseding expression was observed could be seen in the basal cells, and the expression also occureds in the flat cells and wing cells. The maximum predominant expression can be seen at the 3-7 d after the injury. At the 14 d when the epithelium had rehabilitated, the expression decreased in the basal column cells and stromal kerotocytes and disappeared in the flat cells and the wing cells. The expression pattern of HA is the same as CD44. Conclusion CD44 and it's receptor HA play important role in the course of the recovery of corneal alkaline burn. They participate inperform the adhesion of the cellcell and cellmatrix, the epithelial migration and rehabilitation, and can facilitate help the corneal reepithelization.
KEY WORDS: antigens, CD44; eye burns; burns,chemical; hyaluronic acid; disesase models, animal; immunohistochemistry
细胞外基质受体Ⅲ(extracellular matrix receptor type Ⅲ,CD44)是分布极为广泛的细胞表面跨膜糖蛋白,属于未归类的黏附分子,具有多种功能及结构形态,通过与其配体透明质酸(Hyaluronan, HA)的结合而发挥广泛作用。笔者建立角膜碱烧伤大鼠模型,采用免疫组织化学方法观察CD44及HA在角膜碱烧伤后不同时间的表达变化,探讨其在角膜碱烧伤修复过程中的作用和意义,为更有效的治疗角膜碱烧伤提供理论基础。
1 材料和方法
1.1 材料
1.1.1 动物与分组
健康SD大鼠（山东省立医院动物中心提供,符合中心清洁实验级别标准）46只,体质量280～300g,鼠龄6～8周,雌雄各半；随机选取6只作为正常角膜组,10只为角膜机械划伤组,30只为角膜碱烧伤组。
1.1.2 实验试剂
兔抗鼠CD44单克隆抗体,兔抗鼠透明质酸单克隆抗体,即用型SABC免疫组织化学染色试剂盒（中国博士得生物工程公司）；DAB酶底物显色试剂盒,0.1%胃蛋白酶（中国Maxim生物工程公司）。
1.2 方法
1.2.1 角膜碱烧伤大鼠模型制作
参照文献[1]制作大鼠角膜碱烧伤模型：3%戊巴比妥钠0.2 mL腹腔注射麻醉大鼠,取直径为5 mm、浸透0.5 mol/L NaOH的圆形滤纸,放在大鼠角膜中央,30 s后取下,以乳酸林格平衡液1 000 mL冲洗角膜及结膜囊,pH试纸测试为中性。伤后结膜囊涂金霉素眼膏,10丝线缝合眼睑。分别于烧伤后12 h,1,3,7,14 d各随机取出6只杀死,取出角膜。
1.2.2 角膜机械性划伤模型制作
大鼠麻醉后,以角膜环钻钻取大鼠角膜中央上皮及前基质层,直径为5 mm,至裂隙灯下见圆形角膜溃疡灶,结膜囊涂金霉素眼膏防止继发感染,10丝线缝合眼睑。每日观察修复情况并分别于模型建立后12 h,1,3,7,14 d各随机取出2只杀死,取出角膜。
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