维生素A治疗苯扎氯铵诱导的小鼠干眼的实验研究  

Therapeutic effects of Vitamin A on mouse dry eye induced by benzalkonium chloride topical medication

作　　者：何欢[1] 刘祖国[1] 肖辛野[1] 林志荣[1] 周彤[1] 何卉[1] 

机构地区：[1]厦门大学眼科研究所,福建省眼科与视觉科学重点实验室,361005

出　　处：《中华眼视光学与视觉科学杂志》2012年第4期197-202,共6页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：国家自然科学基金资助项目（30872810）

摘　　要：目的通过随机对照实验探讨维生素A对苯扎氯铵诱导的小鼠干眼的治疗作用。方法实验研究。选用BALB＆小鼠50只，使用0．25％的苯扎氯铵溶液局部点眼以诱导干眼。在第21天时根据泪膜破裂时间（BUT）、角膜荧光素钠染色、炎症指数等参数选择干眼表现程度相近的小鼠30只（60眼）并随机分为A、B、C三组，每组10只（20眼）。A、B组分别使用5000IU／（g·mD维生素A微乳剂和空白微乳剂滴眼，C组为空白对照组。在治疗的第0、1、3、5、7天检测BUT和角膜上皮荧光素钠染色、眼表炎症程度及基础泪液分泌量。7d治疗结束后取出小鼠眼球，进行HE及过碘酸．希夫（PAS）染色，角蛋白10（K10）免疫荧光标记，以及WesternBlot检测角膜组织中K10表达水平。对数据采用单因素方差分析。结果在药物治疗的第0、1、3、5天各项临床指标差异无统计学意义。治疗第7天时，A、B、C组BUT值分别为（4．20±0．89）s、（3．32±1．01）s和（3．28±0．74）s；其中A组的BUT值较B组和C组长，差异有统计学意义（F=6．825，P=-0．002）。A、B、C组角膜荧光素钠染色评分分别为3．74±1．31、5．47±1．81和5．83±1．54，其中维生素A微乳组较空白微乳组和空白对照组角膜荧光素钠染色分级轻．差异具有统计学意义（F=8．853，P〈0．01）。各组间的角膜炎症指数评分、基础泪液分泌量差异均无统计学意义。组织病理检查示维生素A组与空白微乳组和空白对照组比较，其角膜上皮形态较规整；PAS染色示维生素A组杯状细胞数量更接近正常小鼠。维生素A组的角膜上皮层几乎均不表达K10．而空白微乳组及空白对照组呈阳性表达。Westem Blot检测示维生素A组角膜中的K10水平低于空白微乳组和空白对照组。结论维生素A可以延长苯扎氯铵诱导的干眼小鼠的BUT，降低角膜上皮染色程度，抑制角膜上皮鳞状化生，�Objective To investigate the therapeutic effect of Vitamin A on the recovery of dry eye that was induced by benzalkonium chloride （BAC） topical medication in the mouse. Methods In this randomized controlled experimental study, 50 BALB/e mice were treated with the topical administration of 0.25% BAC to establish the dry eye condition. Based on the consistency of tear film break-up time （BUT）, corneal fluoreseein staining scores and inflammation index, the eyes were re-selected and 30 mice were randomly divided into three groups on day 21 after the BAC treatment, with 10 mice （20 eyes） in each group. Group C was set up as a blank control, while groups A and B were treated with 5000 IU/（g.ml） Vitamin A eye drops and mieroemulsion solvent drops, respectively. BUTs, corneal fluorescein staining scores, inflammation index and tear volumes were evaluated in each group on days 0, 1, 3, 5 and 7 after the therapeutic treatment. Global specimens were collected on day 7. Sections were stained with hematoxylin and eosin （HE） or by periodic acid-Schiff （PAS） assay, and labeled with cytokeratin 10 （K10） antibody. The expression of K10 in the cornea and conjunctiva was quantified by Western Blot. The data were analyzed by one-way ANOVAs. Results On days 0, 1, 3 and 5, no clinical differences were observed among the groups. On day 7, the BUT values were （4.20±0.89）s, （3.32±1.01）s and （3.28±0.74）s in groups A, B and C, respectively. The BUT in group A was significantly longer than those in groups B and C （F=6.852, P=0.002）. The corneal fluorescein staining scores were 3.74±1.31, 5.47±1.81 and 5.83＋1.54 in groups A, B and C, respectively. The scores in group A were significantly lower than those in groups B and C （F=8.853, P〈0.01）. No significant differences were found in the corneal inflammatory index or tear volume among the groups. Corneal morphology showed a loosened corneal epithelium feature in groups B and C, but corneal integrity in group A. PAS assay revealed similar

关 键 词：干眼病 维生素A 鳞状上皮化生 杯状细胞 模型 动物 小鼠 

分 类 号：R777[医药卫生—眼科]