兔自身免疫性干眼模型的建立及泪腺炎症因子研究  

Establishment of rabbit autoimmune dry eye model and studies of lacrimal cytokines

作　　者：杨丽媛[1] 边领斋[1] 李雪[1] 粘红[1] 东莉洁[1] 魏瑞华[1] 

机构地区：[1]天津医科大学眼科医院,天津医科大学眼科研究所,天津市300384

出　　处：《眼科新进展》2015年第9期810-815,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81100646;31100991);天津市科委自然科学基金资助(编号:11JCYBJC26000;13JCYBJC23300)~~

摘　　要：目的通过静脉回输自体激活的淋巴细胞建立稳定的兔自身免疫性干眼模型,并分析干眼的泪腺炎性因子,探究干眼的泪腺免疫反应。方法通过体外分离、培养兔泪腺上皮细胞和自体外周血淋巴细胞,建立二者的共培养体系。将体外激活的自体外周血淋巴细胞通过耳缘静脉注射的方法诱发自身免疫性泪腺炎。对干眼临床指标及泪腺细胞因子表达进行检测。结果与正常对照组相比,干眼模型组泪液分泌显著减少(t=115.6,P<0.05),泪膜破裂时间缩短(t=92.5,P<0.05)。荧光素钠染色显示角膜弥漫性点状着色,正常对照组基本无着色。泪腺组织HE染色显示淋巴细胞浸润,正常对照组很少。RT-PCR结果显示,干眼模型组泪腺中炎性因子IFN-γ、IL-6、IL-17 mRNA相对表达量显著升高(t=26.2、18.7、8.4;均为P<0.05),抑炎因子IL-10、TGF-βmRNA相对表达量显著降低(t=22.1、14.2;均为P<0.05)。结论经兔泪腺上皮细胞激活的自体外周血淋巴细胞静脉回输后,成功诱发干燥综合征样自身免疫泪腺炎,建立了稳定的兔自身免疫性干眼模型。泪腺组织中细胞因子IL-6、IL-17、IFN-γ、TGF-β、IL-10参与泪腺内炎症反应的发生和发展。Objective To explore the lacrimal immune response and cytokines changes of rabbit autoimmune dry eye model induced by intravenous infusion of autologous activated lymphocytes.Methods The co-culture system of purified rabbit lacrimal gland epithelial cells and autologous peripheral blood lymphocytes was established.Autoimmune dacryoadenitis model was induced by injecting activated autologous peripheral blood lymphocytes into rabbit ear marginal vein.Ocular examinations were performed.The lacrimal gland were removed for histopathology and used for Real-time PCR.Results In dry eye model group,tear secretion significantly decreased（t=115.6,P〈0.05）,break-up time was obviously shortened（t=92.5,P〈0.05）.The sodium fluorescein staining appeared diffuse dot cornea staining compared with the control group.After intravenous injection of activated lymphocytes,HE staining revealed that lacrimal gland had lymphocytic infiltration,but the control group was few.Realtime PCR results revealed that the levels of inflammatory factors such as IFN-γ,IL-6 and IL-17 mRNA from dry eye model group were significantly increased in comparison with control group（t=26.2,18.7,8.4;all P〈0.05）.However,the levels of IL-10 and TGF-β mRNA were significantly decreased in comparison with the control group（t=22.1,14.2,all P〈0.05）.Conclusion Injection of the activated autologous peripheral blood lymphocytes successfully induce Sjogren＇s autoimmune dacryoadenitis.Cytokines IFN-γ,IL-6,IL-10,IL-17 and TGF-β may participate in the inflammatory response,leading to the occurrence of dry eye.

关 键 词：自身免疫性泪腺炎 兔自身免疫性干眼模型 细胞因子 IL-10 

分 类 号：R777.3[医药卫生—眼科]