Th1/Th2细胞因子在小鼠真菌性角膜炎中的表达  

Expression of Th1 and Th2 type cytokines in experimental keratomycosis

作　　者：胡建章[1] 许建斌[1] 黄定国[1] 林秀丽[1] 朱学军[1] 韩晓丽[1] 徐国兴[1] 

机构地区：[1]福建医科大学附属第一医院眼科福建省眼科研究所,福州350005

出　　处：《中国实用眼科杂志》2010年第8期919-922,共4页Chinese Journal of Practical Ophthalmology

基　　金：福建省科技厅青年人才项目（2008F3041）,福建省卫生厅青年科研基金项目（2007-2-1;2008-2-18）,福建医科大学教授发展基金（2006js6033;js09009）

摘　　要：目的 通过检测Th1/Th2细胞因子在小鼠真菌性角膜炎中的表达水平,探讨机体免疫在该病发展中的作用. 方法应用角膜表层镜法建立Balb/c小鼠茄病镰刀菌性角膜炎模型,在感染后第1、3、5、7天,裂隙灯显微镜观察角膜炎特点;HE染色观察角膜病理变化;半定量RT-PCR和ELISA检测角膜中Thl细胞因子（IFN-γL-12）及Th2细胞因子（IL-4、IL-10）基因mRNA和蛋白的表达,半定量RT-PCR检测T-bet基因mRNA的表达;直线相关分析检测T-bet mRNA与IFN-比值的相关性. 结果角膜接种菌液后,早期角膜浸润混浊进行性加重,第5天后新生血管大量生长;HE染色可见第1、3天在角膜缘、角膜基质及前房中有大量的炎症细胞浸润,第5天后炎症细胞减少,角膜基质中纤维细胞逐渐增多;RT-PCR与ELISA检测结果表明,Thl细胞因子（IFN-γL-12）和Th2细胞因子（IL-4、IL-10）基因mRNA及蛋白在角膜接种菌液后均出现表达,且IFN-γL-12的表达显著强于IL-4、IL-10;IFN-γ/IL-4比值在第3天达最高值,而后逐渐降低;T-bet mRNA在第3天达最高值;T-bet mRNA的表达与IFN-γ/IL-4比值呈正相关（P＜0.05）. 结论在真菌性角膜炎中,Thl/Th2型免疫应答共同参与调节机体的抗真菌免疫,但以Th1型应答为主;角膜感染真菌后第3天机体的免疫力达最强.Objective To detect the expression of Th1 and Th2 type cytokines in a mouse model of Fusarium solani keratitis, and investigate the role of immune response in this disease. Methods A mouse model of Fusarium solani keratitis was established by epikeratophakia. On day 1, 3, 5, 7 after inoculation, the appearance of corneal ulcer was observed and infected corneas were subjected to histopathological analysis. Th1 （IFN-γ, IL-12） and Th2 （IL-4, IL-10） cytokines production in the cornea was analyzed by enzyme-linked immunosorbent assay （ELISA）, and cytokine mRNA levels were measured by semiquantitative reverse transcription-polymerase chain reaction （RT-PCR）. T-bet mRNA level were detect by RT-PCR. Results The infected cornea was characterized by severe inflammation, diffuse edema and opacity, and much neovascularization occurred on 5 days after inoculation. Histopathologic examination showed at 1 day and 3 days after inoculation, the limbus, corneas and anterior chamber had marked dense inflammatory cell infiltration, extensive stromal collagen destruction and edema. However, at 5 and 7 days, with a decrease of PMNs infiltration, significant neovascularization and uneven thickening and much fibrocytes could be seen in the stroma. Both Thl and Th2 cytokine production and mRNA expression in the corneas were detected, and the Th1 cytokine levels were much higher than Th2 cytokine. The ratio of IFN-γ/IL-4 reached to the peak at 3 days, then decreased gradually. There was positive correlation between T-bet mRNA expression and the ratio of IFN-γ/IL-4 （P 〈0.05）. Conclusions In fungal keratitis, both the Th1 and Th2 responses are involved in regulating the anti-fungal immunity, but the Th1 response is dominant in the course. The power of anti-fungal immunity is rised to peak on day 3 after inoculation.

关 键 词：真菌性角膜炎 TH1/TH2细胞因子 免疫应答 

分 类 号：R512.62[医药卫生—内科学]