20%乙醇处理兔角膜后上皮增生和细胞凋亡的研究  

Proliferation of corneal epithelium and apoptosis of keratocytes in the rabbit cornea after treatment with 20% ethanol

作　　者：孙丽霞[1] 王铮[1] 杨斌[1] 刘娟[1] 邱平[1] 陈家祺[1] 

机构地区：[1]中山大学中山眼科中心,广州510060

出　　处：《中华眼科杂志》2005年第6期492-497,共6页Chinese Journal of Ophthalmology

基　　金：广东省科技计划基金资助项目(2002C30902)

摘　　要：目的探讨准分子激光角膜上皮瓣下磨镶术(LASEK)中采用20%乙醇浸润兔角膜40s后角膜上皮增生和角膜细胞凋亡情况与机械刮除角膜上皮后的异同。方法实验组42只新西兰大白兔,用直径为8mm的LASEK专用角膜上皮刀切割角膜上皮,20%的乙醇浸润单眼40s,机械刮除对侧眼中央8mm直径的角膜上皮,随机分7组,于术后0、4h,1、3、5、8、30d取材;6只兔眼为空白对照。角膜冰冻切片,行Ki67免疫组化检查和TUNEL检测,计数角膜中央前基质细胞。结果乙醇浸润后5d中央角膜上皮增生达峰值,术后1d周边角膜上皮增生达峰值;术后4h上皮刀口下方局限的角膜基质细胞TUNEL染色阳性,数量最多;各组角膜中央前基质细胞计数和空白对照比差异无统计学意义(P=0.68)。机械刮除角膜上皮后3d周边角膜上皮增生达峰值,其高于乙醇浸润后角膜上皮的增生峰值;术后4h可见大量中央前基质细胞TUNEL阳性;术后1d中央前基质细胞数量最少(P<0.05)。结论与机械刮除角膜上皮相比,20%乙醇浸润40s对角膜损伤轻,恢复快,乙醇浸润后的角膜上皮对基质细胞有保护作用。Objective To study the proliferation activity of corneal epithelial cells and the apoptosis of keratocytes in the rabbit cornea after treatment with 20% ethanol. The results were compared to the cornea treated with mechanical scraping of the epithelial cells. Methods The experimental group consisted of 42 rabbits. One of the two corneas of each rabbit was incised by 8 mm epithelial trephine using in LASEK and was exposed to 20% ethanol (in distilled water) in the trephine for 40 seconds. In the other eye, the corneal epithelium in the central area was mechanically scraped. The rabbits in the experimental group were randomly divided into seven sub-groups. Each sub-group had six rabbits and the rabbits were killed at 0, 4 hours, 1, 3, 5, 8 and 30 days after the surgery. Three rabbits without any treatment were used as blank controls. Immunohistochemical staining (Ki-67 antigen) was performed to detect the proliferation of corneal epithelial cells. Apoptotic cells were detected by TUNEL assay. Number of keratocytes in the central anterior stroma of cornea was determined by counting the total number of cells under 400×magnification field in hematoxylin-eosin-stained corneal sections. Results In the ethanol-treated eyes, the number of Ki-67 positive cells peaked 5 days after the treatment in the central corneal epithelium and 1 day after the treatment in the peripheral corneal epithelium. TUNEL-positive cells were detected in the anterior central stromal keratocytes under the epithelial incisions and the number of TUNEL-positive cells reached a peak 4 hours after the treatment. There was no statistically significant difference in the number of central anterior stromal keratocytes between the ethanol-treated group and the controls (P=0.68). In the mechanical scraping group, the number of Ki-67 positive cells in the peripheral corneal epithelium peaked 3 days after the treatment . The number of Ki-67 positive cells in the peripheral corneal epithelium of the scraping group was greater than that of the ethanol-treated g

关 键 词：上皮增生 准分子激光角膜上皮瓣下磨镶术 角膜后 醇处理 TUNEL染色 机械刮除 角膜细胞凋亡 新西兰大白兔 免疫组化检查 角膜基质细胞 空白对照 角膜中央 LASEK Ki-67 冰冻切片 中央角膜 细胞计数 细胞数量 角膜损伤 保护作用 

分 类 号：R779.63[医药卫生—眼科]