实验性自身免疫性葡萄膜炎炎性因子表达的动态观察  

Dynamic expression of inflammatory factors in experimental autoimmune uveitis in mice

作　　者：王影[1] 李洋[2] 毕宏生[2] 滕达[1] 李姣[1] 崔彦[2] 

机构地区：[1]山东中医药大学,济南250002 [2]山东中医药大学附属眼科医院山东中医药大学眼科研究所,济南250002

出　　处：《中华实验眼科杂志》2013年第7期647-652,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81072961、81100658）;山东省自然科学基金项目（ZR2010HM048）

摘　　要：背景C57BL／6（B6）小鼠是实验性自身免疫性葡萄膜炎（EAU）动物模型常用的小鼠种系，以往研究证明葡萄膜炎的发病机制与辅助性T（Th）细胞分泌的炎性细胞因子相关，但各种Th细胞在EAU发病中的相互作用并不十分清楚。目的研究光感受器间维生素A类结合蛋白（1RBP）诱导不同天数后EAU小鼠脾脏和血清中炎性因子的动态变化，探讨各种炎性因子在EAU病理过程中的作用。方法用IRBP及完全弗氏佐剂（CFA）的乳化液在小鼠的尾根部及躯干部均匀注射5个点以免疫B6小鼠44只，免疫后每周3次用间接检眼镜观察小鼠的EAU发病情况，并参照Thurau的评分标准进行炎症评分。于注射后第30天摘取20只模型小鼠眼球，于瞳孔视神经平面制备切片行组织病理学检测。分别于注射前，注射后第2、5、10、15、20、25、30天取模型小鼠脾脏，提取RNA，逆转录扩增并行凝胶电泳，检测白细胞介素-17（IL-17）、叮干扰素（IFN-γ）、肿瘤坏死因子-α（TNF—α）和IL-10等因子的mRNA含量，同时收集相同时间点小鼠的血清，采用酶联免疫吸附试验（ELISA）检测血清中上述因子的质量浓度。结果IRBP及CFA乳化液免疫B6小鼠后第12天可见轻度葡萄膜炎症，炎症评分为0．5分，炎症反应在免疫后第13～15天最重，评分为1．0分，至第30天炎症明显减轻，评分为0．5。组织病理学检查显示，注射后第30天模型鼠眼部反应与间接检眼镜检查结果吻合，病理评分为0．5。模型鼠血清炎性因子检测结果显示，注射后第5天，小鼠血清中IL-17的质量浓度达到峰值，为（51．85±2．42）ng／L，随着时间的延长开始降低。到第15天时降到最低，为（4．01±0.06）ng／L，但在第25天时再次升高至（25．00±0．94）ng／L，之后逐渐下降，第30天时，血清中IL-17的质量浓度为（6．01±0．21）ng／L，与免疫前的（0．98±0．05）ng�Background C57BL/6（B6） is a kind of routine mouse specie used in experimental autoimmune uveitis （EAU） research. Previous studies showed that the pathogenesis of uveitis related to inflammatory cytokines secreted by different helper T （Th） cefls. However, the interaction of different Th cells in EAU is unclear. Objective This study was to investigate the dynamic changes of inflammatory factors in the spleen and serum after immunization in EAU mice. Methods Forty-four SPF B6 mice were immunized by injection of interphotoreceptor retinoid-binding protein （IRBP） and complete Freund adjuvant （CFA） emulsion via caudal vein and footpad. Indirect ophthalmoscope was used to examine the eyes 3 times per week and the inflammatory response was scored based on Thurau＇s criteria. In the thirty day after injection, 20 model eyes were extracted and the sections of eye tissue were prepared for histopathological examination. The spleens of model mice were enucleated before injection and 2, 5, 10, 15, 20, 25,30 days after injection, and reverse transcriptase PCR （RT-PCR） was used to detect the contents of interleukin-17 （IL-17） mRNA, interferon-3, （IFN--γ） mRNA, tumor necrosis factor-or （TNF-ot） mRNA and IL-10 mRNA, and the contents of IL-17, IFN-±,, TNF-±x and IL-10 in model serum were assayed by ELISA in 24 model mice. The experimental protocol and use of the animals were approved by Ethic Committee for Care and Use of Laboratory Animals of Shandong University of Traditional Chinese Medicine. Results Mild inflammatory response was seen in 12 days under the indirect ophthalmoscope with the scores of 0.5. The inflammatory scores peaked in 13- 15 days with the scores of 1.0 and alleviated after that with the inflammatory scores of 0.5 in 30 days after injection. The histopathological score was consistent with the clinical score in the models on the 30 days. The serum IL-17 content of model mice was （0.98 ±0. 05 ） ng/L before injection and increased to （ 51.85±2.42） ng/L on the fift

关 键 词：葡萄膜炎 实验性自身免疫性 免疫 特异性自身免疫疾病 辅助T细胞 炎性因子 白细胞介素 γ干扰素 肿瘤坏死因子-α C57BL 6小鼠 

分 类 号：R773.9[医药卫生—眼科]