眼科研究 2000年第3期第18卷 实验研究

作者：谢雷克　张俊杰　陈祖基　徐岩

单位：河南省科研究所，郑州450003

关键词：酮替芬；微量激素；过敏性结膜炎

　　摘要　目的　观察复方酮替芬滴眼剂(含0.1%酮替芬及0.0005％地塞米松)对实验大鼠急性过敏性结膜炎的治疗作用。　方法　雄性Wistar大鼠27只随机分为3组(复方酮替芬、0.1%酮替芬及地塞米松组)，每组9只，各预先腹腔注射卵清蛋白致敏液（100μg卵清蛋白及20mg硫酸铝钾溶于1ml磷酸盐缓冲液，pH7.4)，两周后以10%卵清蛋白液(溶于磷酸盐缓冲液)10μl滴眼攻击，引起结膜速发型过敏反应。攻击前15min以1mol^．l^－1DL-二巯苏糖醇20μl作眼部预处理，临攻击时静脉注射Evans蓝(EB)溶液(约2mg／100g）；攻击前60，45，30，15min及攻击后15，30min各组1眼给药，对侧眼为溶媒。攻击后1h处死所有动物，测定各眼EB渗出量，进行比较，计算药物抑制率。　结果　各组药物治疗眼EB外渗量均低于溶媒治疗眼，复方酮替芬组和0.1%酮替芬组显示统计学有显著意义。复方酮替芬抑制率为53.98％±15.57％，显著高于0.1%酮替芬（27.91％±28.36％，P＝0.028）及0.0005％地塞米松（11.90％±34.16％，P＝0.004）。　结论　结果提示复方酮替芬滴眼剂具有良好的抗过敏作用。

　　分类号　R777.3

Treatment of experimental allergic conjunctivitis with compound ketotifen eye drops

Xie Leike，Zhang Junjie，Chen Zuji，Xu Yan

　　（Henan Institute of Ophthalmology，Zhengzhou 450003，China）

　　Abstract　Objective：To examine the effect of compound ketotifen eye drops（CKF，containing 0.1％ ketotifen and 0.000 5％ dexamethasone） on experimental acute allergic conjunctivitis in rats．Methods：27 male Wistar rats were immunized by intraperitoneal injection of egg albumin and were randomized into three groups： CKF，0.1％ ketotifen （KF） and 0.000 5％ dexamethasone （DM） Rats in each group received the respective eye drops in one eye and vehicle in the other eye 60，45，30，15 min before and 15，30 min after challenge．Immediately before challenge，the rats were injected intravenously with Evans Blue（EB）．The challenge was performed by topical instillation of egg albumin，and 60 min later，the animals were killed and the dye was extracted from the eyes．The intensity of EB extravasation was determined by spectrophotometry at 620nm and inhibitory rates were calculated．Results：CKF，KF and DM suppressed EB extravasation significantly．The inhibitory rate was 53.98％±15.57％ for CKF，27.91％±28.36％ for KF and 11.90％±34.16％ for DM．CKF was more effective than both KF and DM on inhibiting dye vascular leakage in the eyes （P＝0.028，0.004，respectively）．Conclusion：These data clearly indicate that the compound of ketotifen eye drops has potential as a topical ocular formulation for anti-allergic disease．

　　Key words　ketotifen micro-corticosteroid allergic conjunctivitis

　　Ketotifen is a potential anti-allergic agent．It has been showed that ketotifen was greatly effective and well tolerated for clinical treatments of seasonal and perennial rhinitis，bronchial asthma and atopic dermatitis^［1］．In the present study，we evaluated an effectiveness of compound ketotifen eye drops prepared with ketotifen （0.1％） and micro-dexamethasone（0.0005％） on experimental ocular immediate hypersensitivity as compared to each drug administered alone．

　　We used a model of topically induced ocular anaphylaxis in rats，which were immunized and challenged with egg albumin（EA） in the study．This ocular model was quite similar to the human conjunctival allergic disease．The severity of the allergic reaction and efficacy of anti-allergic drugs were assessed objectively by quantifying evans blue（EB） extravasation in ocular tissue．

　　MATERIALS %26 METHODS

　　Animals Twenty seven male Wistar rats，weighing 250～300g，were purchased from Henan Experimental Animal Center．

　　Drugs and reagents Ketotifen（Shanghai），Dexamethasone（Shanghai），DL-Dithiothreitol （Shanghai），Egg Albumin （Sigma），and Evans Blue（TCI，Japan） were used．Solutions for making eye drops，including compound ketotifen （CKF），0.1％ ketotifen （KF），and 0.0005％ dexamethasone（DM），were prepared using the same vehicle．

　　Instruments Spectrophotometer UV／V 7530-G（Shanghai，China）and electronic analytical balance AE 260（Mettler，Switzerland）were used．

　　Immunization and challenge All rats were injected intraperitoneally with 100μg EA plus 20mg of alum in 1ml of phosphate buffered saline（PBS），pH7.4 for immunization.14d later，rats were challenged topically with 10μl EA 10％ in PBS．To obtain a remarkable ocular reaction，20μl of 1mol／L DL-Dithiothreitol，a mucolytic agent，in PBS（freshly prepared before use）was applied to each eye 15min before topical challenge as described by Calonge et al^［2］ and Trocme et al^［3］．EB（2mg／100g） in 1ml of PBS was intravenously injected immediately prior to challenge．Animals were anesthetized with intraperitoneal 5％ chloral hydrate（about 0.2ml／100g）．

　　Drug administration Animals were divided into three groups（CKF，KF and DM）of nine each．One eye of each animal received one drop of relevant drug and the fellow eye vehicle 60，45，30，15min before and 15，30min after challenge．

　　Evaluation procedure All rats were killed by exsanguination 1h after challenge．Globes and adnexa were exenterated，weighed，and then immersed in an extracting solution of 5ml sodium sulfate（0.5％） plus acetone（3∶7，v／v），shaken strongly，and kept at room temperature．24h later，solutions were centrifuged at 1000rpm for 10min．The color intensity of supernatant was detected spectrophotometrially at 620nm．

　　Standard curve was performed to transform absorbance unit in microgram of EB per milliliter of solution （μg／ml） and EB extravasation in ocular tissues was calculated．Drug efficacy were expressed as percent inhibition of the reaction on the vehicle treated eye．IR（％）＝（CV－CD）／CV；IR：drug inhibitory rate；CD is EB content in drug treated eye and CV in the untreated control eye．Data were analyzed by paired and unpaired，two-tailed student t-test．A P value of 0.05 or less was considered significant．

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