摘要
目的观察胰岛素对兔角膜碱烧伤后角膜组织病理学的变化以及对白细胞介素(IL)表达的影响,并探讨胰岛素在角膜碱烧伤后早期的抗炎机制。方法选取45只健康雌性新西兰白兔,随机分为生理盐水组、地塞米松组和胰岛素组,每组15只。应用浸有1mol/LNaOH溶液的滤纸贴附于角膜中央建立Ⅱ°角膜碱烧伤模型,建模后即日起各组分别于球结膜下注射生理盐水、地塞米松和胰岛素各75μL/(kg.d),于实验后1、3、7d收集每只动物心脏血3mL和每只眼200μL房水,分别用ELISA法检测实验动物血清和房水中IL-1β的质量浓度的变化,应用逆转录聚合酶链反应(RT-PCR)法检测角膜组织中IL-10mRNA浓度的变化,并对角膜组织进行组织病理学检查,观察角膜组织的形态变化。结果角膜碱烧伤后第1天生理盐水组、地塞米松组和胰岛素组均可见角膜上皮缺失、基质层严重水肿和角膜组织大量炎性细胞浸润;碱烧伤后第3天3组角膜上皮和基质损伤均有不同程度的修复,至第7天生理盐水组角膜基质仍水肿,但地塞米松组和胰岛素组角膜基质层水肿基本消退。角膜碱烧伤后第1、3、7天,3组实验兔血清和房水中的IL-1β质量浓度明显高于正常兔,但地塞米松组和胰岛素组实验兔血清和房水中的IL-1β质量浓度明显低于生理盐水组,差异均有统计学意义(P<0.01),第3天、第7天地塞米松组和胰岛素组血清和房水中的IL-1β质量浓度的变化差异均无统计学意义(P>0.05)。角膜碱烧伤后各时间点地塞米松组和胰岛素组角膜组织中IL-10mRNA浓度明显高于生理盐水组,差异均有统计学意义(P<0.01),但地塞米松组和胰岛素组间的差异无统计学意义(P>0.05),实验后第3天、第7天地塞米松组和胰岛素组角膜组织中IL-10mRNA浓度均明显高于第1天,差异均有统计学意义(P<0.01)。结论角膜碱烧伤早期胰岛素可以通过减少促炎性因子的表达、提高抑制炎性因子含量的机制起到抗炎作用,促进角膜损伤的修复。胰岛素的抗炎作用类似于地塞米松。
Background Corneal alkali burn is usually lend to blindness,and the pathological changes is closely related to inflammation.Recent research demonstrated the anti-inflammatory effect of insulin.ObjectiveThis study aimed to observe the effects of insulin on corneal histopathological change and interleukin expression after corneal alkali burn in rabbit and explore the early anti-inflammatory mechanism of insulin.MethodsForty-five female New Zealand white rabbits were randomly divided into saline group,dexamethasone group and insulin group and 15 rabbits for each group.Corneal alkali burn models established by putting the 1 mol/L NaOH filter paper at the central cornea for 40 seconds.75μL/kg of normal saline solution,dexamethasone and insulin were subconjunctically injected once per day according to the grouping.Other 5 normal rabbits were used as normal control group.3 mL blood and 200μL aqueous humor were collected in 1,3,7 days after operation.Concentration of interleukin-1β（IL-1β）in blood and aqueous humor were detected by ELISA.The corneal tissue section was prepared in the same time points for histopathological examination.Content of IL-10 mRNA in corneal tissue was detected by reverse transcription-PCR.Results Deficiency of corneal epithelium,edema of corneal stroma and infiltration of inflammatory cells were found in normal saline solution,dexamethasone and insulin groups in the first day after operation.On the seventh day,cornea edema was still existed in normal saline group and almost disappeared in dexamethasone and insulin groups.The concentration of IL-1β in blood and aqueous humor was significantly elevated in 1,3,7 days after operation in 3 groups compared with normal rabbits,but that of dexamethasone and insulin groups was lower than the normal saline group（P0.01）.No evident difference was found in the concentration of IL-1β between dexamethasone group and insulin group in 3 and 7 days after operation（P0.05）.The IL-10 mRNA level in corneal tissue in dexamethasone and insulin groups were significantly increased than that in normal saline group at 1,3 or 7 day respectively（P0.01）,but no evident difference was found in IL-10 mRNA level between dexamethasone group and insulin group.Compared with the first day,IL-10 mRNA level in corneal tissue was evidently increased in the third and seventh day after operation in dexamethasone group and insulin group（P0.01）.ConclusionInsulin play an anti-inflammation action and promotion effect of restoration of corneal injury by lowing the pro-inflammatory factors and improving the content of anti-inflammatory factors in the early stage of corneal alkali burn.
出处
《眼科研究》
CSCD
北大核心
2010年第11期1036-1041,共6页
Chinese Ophthalmic Research
