摘要
目的探讨普拉洛芬对苯扎氯铵诱导小鼠干眼的治疗作用及其可能机制。方法实验研究。选用BALB／c雄性小鼠70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似的眼并随机分为A、B、C、D四组。A组为空白对照，B、C、D组分别使用0．1％玻璃酸钠滴眼液，0．1％玻璃酸钠滴眼液加0．1％氟米龙滴眼液，0．1％玻璃酸钠滴眼液加0．1％普拉洛芬滴眼液进行治疗。治疗后第0、1、3、5天进行BUT检测、角膜上皮荧光素钠染色，眼表炎症程度的评估及基础泪液量的测量。治疗结束后取下小鼠眼球，进行HE及过碘酸一希夫（PAS）染色，角蛋白10（K10）免疫荧光标记，以及Westernblot检测角结膜组织中肿瘤坏死因子α（TNF—α）表达水平。对方差齐性的数据采用Tukey显著性检验，对方差非齐性的数据采用Dunnett检验。结果经筛选共36只鼠（72只眼）人选治疗实验，每组18只眼。在使用药物治疗后的第0、1、3天各项临床指标差异无统计学意义。治疗第5天时，A、B、c、D组泪BUT分别为（2．933±0．320）、（2．900±O．280）、（3．464±0．498）和（3．643±0．413）S；其中C组与D组的BUT均较A组和B组延长，差异具有统计学意义（F=13．774，P=0．000）。角膜荧光素钠染色分级A、B、C、D组分别为（11．640±1．008）、（11．790±1．188）、（10．330±1．371）及（10．270±1．104）分；两联合用药治疗组均较人工泪液组和空白对照组角膜荧光素钠染色分级减轻，具有统计学意义（F=6．145，P=0．001）。角膜炎症指数评分A、B、C、D组分别为（0．232±0．059）、（0．229±0．078）、（0．151±0．055）及（0．154±0．056）分；两联合用药治疗组均较人T泪液组和空白对照组炎症指数降低，差异具有统计学意义（F=6．703，P=0．001）。各组间的基础泪液分泌量差异无统计学意义。HE结果显示空白对照组上皮厚度高于两联合用药组，联合用药组上皮规整；PAS染色示普拉洛芬组及氟米龙组的杯状细胞数量相当，均多于人工泪液组及空白对照组。普拉洛芬组及氟米龙组的角膜上皮层均几乎不表达K10，而人工泪液组及空白对照组仍阳性表达。普拉洛芬组及氟米龙处理组角膜中的TNF-α水平均有下降。结论普拉洛芬具有抑制苯扎氯铵溶液局部点眼诱导干眼眼表炎症的作用，有望用于干眼眼表炎症的临床治疗。
Objective Randomized controlled experimental study to investigate the therapeutic effect and the possible mechanism of Pranoprofen on the recovery of dry eye induced by topical medication of Benzalkonium Chloride （BAC） in mouse. Methods It was an experimental study. Seventy BALB/c mice were treated with topical administration of 0. 25% BAC to establish the dry eye condition. Based on the consistency of break-up time of tear-film （BUT）, corneal fluorescein staining scores and inflammation index, the eyes were re-selected and randomly divided into four groups on day （D） 21 after the BAC treatment. Group A was set up as blank control, while group B, C and D were treated respectively with 0. 1% sodium hyaluronate eye drops, 0. 1% fluorometholone eye drops plus 0. 1% sodium hyaluronate eye drops, 0. 1% pranoprofen eye drops plus 0. 1% sodium hyaluronate eye drops. BUTs, tear volumes, corneal fluorescein staining scores and inflammation index were evaluated in each group on DO, 1, 3 and 5 after the therapeutic treatment. Global specimens were collected on D6. 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 tumor necrosis factor-α （TNF-α） in the cornea and conjunctiva was quantified by western blot. Results 72 eyes were included in the sequential experiment, 18 eyes for each group. On DO, 1 and 3, no clinical differences were observed among the groups. On D5, the BUT was （ 2. 933 ± 0. 320 ） , （ 2. 900 ± 0. 280） , （3. 464 ±0. 498） and （3. 643 ±0. 413）s in group A, B, C and D respectively; the BUTs in group C and D were significant longer than those of group A and B （ F = 13. 774, P = 0. 000 ）. The corneal fluorescein staining score was （ 11. 640 ± 1. 008）, （ 11. 790 ± 1. 188）, （ 10. 330 ±1. 371 ） and （ 10. 270 ± 1. 104）s in group A, B, C and D respectively; the scores in group C and D were significant lower than those of group A and B （F =6. 145 ,P =0. 001 ）. The corneal inflammatory index was （0. 232 ±0. 059）, （0. 229 ±0.078）, （0. 151 ±0.055） and （0. 154 ±0.056） in group A, B, C and D respectively; the index in group C and D were significant lower than those of group A and B （F =6. 703 ,P =0. 001 ）. No significant difference was found in tear volume among groups. No significant difference was found between Pranoprofen and Fluorometholone treatment in BUT, corneal fluorescein score or inflammatory index. Corneal morphology showed the feature of thicker corneal epithelial layer in group A and uniformity in group C and D. PAS assay revealed similar goblet cell numbers in group C and D, but less goblet cells in group A and B. Cytokeratin 10 was almost negatively expressed in Pranoprofen or Fluorometholone treated groups, and remained positive in the corneal epithelium with other treatments. The level of TNF-α in the cornea was down-regulated in Pranoprofen or Fluorometholone treated groups. Conclusions Pranoprofen or Fluorometholone combined with sodium hyaluronate treatment presented similar therapeutic effects on BAC-indueed mouse dry eye, with the more stable tear film, the better regularity of epithelium recovery, the down-regulation of inflammatory TNF-α, the increased number of goblet cells, and the elimination of squamous metaplasia, when compared with the treatment of sodium hyaluronate eye drops only. Our results showed the great potentialities of Pranoprofen in the clinical treatment of ocular surface inflammation in the mild and severity dry eye.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2012年第1期33-40,共8页
Chinese Journal of Ophthalmology
基金
国家自然科学基金（NSFC NO.30872810）
关键词
干眼病
消炎药
非甾类
丙酸类
苯吡喃类
小鼠
肿瘤坏死因子A
Xerophthalmia
Anti-inflammatory agents, non-steroidal
Benzopyrans
Propionic acids
Mice
Tumor necrosis factor-alpha
