重组胸腺素β4在干眼模型中对炎症反应的调节作用及对眼表修复的影响  

Promoting ocular surface restoration effects and anti-inflammatory mechanism of thymosin beta 4 in rat dry

作　　者：张慧芹[1] 李颖[1] 冯丽娜[1] 许永根[1] 

机构地区：[1]北京大学第三医院眼科,100191

出　　处：《中华实验眼科杂志》2017年第2期114-121,共8页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景研究证实炎症与干眼的发生和发展相关，其中肿瘤坏死因子-α（TNF-α）和γ干扰素（IFN-γ）是重要的炎症因子。胸腺素β4（Tβ4）具有促进上皮细胞迁移和抑制炎症反应的作用，但其对干眼眼表修复的影响及其机制尚未阐明。 目的观察重组Tβ4对干眼模型鼠眼中TNF-α和IFN-γ表达的调控作用及其对眼表修复的影响。 方法选用清洁级雄性成年SD大鼠50只，用质量分数0.3%苯扎氯铵溶液连续点左眼7 d以诱导干眼模型，以泪膜破裂时间（BUT）、角膜荧光素染色评分、Schirmer试验Ⅰ（SⅠt）结果评估造模情况。将造模成功的36只眼按随机数字表法分为模型对照组、重组人表皮生长因子（rhEGF）点眼组、Tβ4点眼组及PBS点眼组，按照分组分别用5 μl Tβ4溶液（9 μg/ml）、rhEGF滴眼液、无菌PBS点眼，每日3次，连续7 d，大鼠右眼不进行任何干预作为正常对照组。各组大鼠均于点眼后7 d行BUT、角膜上皮荧光素染色和SⅠt检查。点眼后7 d以过量麻醉法处死动物，制备大鼠眼表组织切片，采用苏木精－伊红染色法观察各组大鼠角膜和结膜组织形态学变化；采用过碘酸希夫染色法计数结膜组织中杯状细胞的数目；透射电子显微镜下观察大鼠角膜和结膜细胞超微结构改变；分别采用实时荧光定量PCR法和Western blot法检测大鼠结膜组织中TNF-α mRNA和IFN-γ mRNA及其蛋白的表达。 结果正常对照组、模型对照组、rhEGF点眼组、Tβ4点眼组和PBS点眼组大鼠BUT分别为（10.42±0.66）、（7.46±0.49）、（8.71±0.50）、（9.59±0.35）和（8.63±0.68）s，总体比较差异有统计学意义（F＝5.65，P＝0.00），其中模型对照组大鼠BUT明显短于正常对照组，rhEGF点眼组和Tβ4点眼组大鼠BUT明显较模型对照组大鼠延长，差异均有统计学意义（均P〈0.05）。各组间大鼠SⅠt和角膜荧光素染色评分的总体比较差异�BackgroundStudies showed that inflammation is associated with the pathogenesis and development of dry eyes, and tumor necrosis factor-α （TNF-α） and interferon-γ （IFN-γ） are key inflammatory factors.Thymosin β4 （Tβ4） plays a promoting effect on the migration of epithelial cells and anti-inflammatory action.However, the influences of Tβ4 on the repair of ocular surface in dry eyes are unelucidated. ObjectiveThis study was to investigate the regulation of Tβ4 to the expressions of TNF-α and IFN-γ and its effect on the recovery of ocular surface in rat dry eye models. MethodsThe dry eye models were induced by topically administered of benzalkonium chloride （BAC） for consecutive 7 days in the left eyes of 50 SPF male SD rats, and 36 successful models were used in the experiment.Tβ4 solution （9 μg/ml）, recombinant human epithelial growth factor （rhEGF） and sterile PBS at 5 μl was topically administered three times for consecutive 7 days in the Tβ4 group, rhEGF group and PBS group, and no drug was used in the model control group.The normal right eyes of rats served as the normal control group.The break-up time of tear film （BUT）, corneal fluorescein staining score and Schirmer Ⅰ test （SⅠt） were examined and evaluated in the rats on the seventh day after administration of drugs.Then the rats were sacrificed by excessive anesthesia and the sections of the ocular surface were prepared.The morphology of the specimens was examined by hematoxylin and eosin staining, and the number of conjunctival gobelt cells was counted by periodic acid-schiff staining.The ultrastructure of the corneal and conjunctival cells was examined under the transmission electron microscope.The expressions of TNF-α mRNA and IFN-γ mRNA and their proteins in conjunctiva tissue were quantified by quantitative real-time PCR and Western blot, respectively.The use and care of the animals followed by Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Techno

关 键 词：干眼/化学诱导 胸腺素β4/治疗作用 眼表 修复 炎症因子 疾病模型 SD大鼠 

分 类 号：R777.34[医药卫生—眼科]