白细胞介素-9在单纯疱疹病毒性角膜基质炎小鼠角膜组织中的表达  

Expression of interleukin-9 in murine cornea with herpetic stroma! keratitis

作　　者：孙超[1] 夏丽坤[1] 

机构地区：[1]中国医科大学附属盛京医院眼科,沈阳110004

出　　处：《中华实验眼科杂志》2013年第2期122-126,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（30772394）;沈阳市高技术产业发展项目计划（2011-14）

摘　　要：背景单纯疱疹病毒性角膜基质炎（HSK）是一种由CD4＋T淋巴细胞介导的免疫病理性疾病。国外研究发现，白细胞介素一9（IL一9）在多种免疫性疾病的发病过程中发挥重要作用，但在HSK中是否起作用目前少有报道。目的研究小鼠HSK发生和发展过程中IL一9在角膜组织中的表达，探讨IL一9的表达与HSK进展程度之间的关系。方法4～6周龄清洁级健康BALB／c小鼠180只分为对照组20只和实验组160只。对照组仅进行“#”字形划痕，不接种病毒；实验组小鼠角膜用注射针划痕后接种5μl含有1×10^6空斑单位（PFU）／ml的I型单纯疱疹病毒（HSV-1）KOS毒株建立HSK小鼠模型。分别于病毒接种前（0d），接种后1、3、5、7、8、10、14、21d在裂隙灯显微镜下观察小鼠角膜的变化；于接种后14d制备角膜组织切片，苏木精一伊红染色观察角膜的组织病理学改变；采用荧光实时定量PCR（real-timePCR）法检测IL-9mRNA在小鼠角膜组织中的表达；采用免疫荧光组织化学方法检测IL-9蛋白在小鼠角膜组织中的表达。结果裂隙灯显微镜下观察结果显示，小鼠角膜接种HSV-1后第3天，实验组小鼠出现点状、树枝状、地图状角膜上皮缺损，接种后6d内上皮缺损痊愈，但于接种后第7天起出现角膜基质灰白色混浊，14d基质混浊加重并达高峰，之后逐渐减轻。Real—timePCR结果显示，未接种病毒（0d）的正常小鼠角膜组织中仅有极微量的IL-9mRNA表达，实验组小鼠接种病毒后1、3、5、7、8、10、14dIL-9mRNA在角膜组织中的相对表达量分别为6．37±0．45、5．66±0．53、3．93±0．35、5．62±0．34、3．23±0．18、2．57±0．14、2．19±0．20，总体表达差异有统计学意义（F=92．764，P=0．000），其中接种后各时间点IL-9mRNA相对表达量与0d比较明显增高，差异均有统计学意义（P〈0．05），接种病毒后21d，IL-9mRNA相对表达量与0d比�Background Herpetic stromal keratitis （HSK） is an immunopathologic eye disorder mediated by CD4~ T lymphocytes. Interleukin-9 （IL-9） is a cytokine linked to the process of many immune inflammatory diseases,but whether IL-9 is involved in the immunopathology of HSK remains unclear. Objective This study was to investigate the expression of IL-9 in murine cornea during the development of HSK and its relationship with the degree of HSK. Methods One hundred and eighty clean BALB/c mice were divided into the normal control group （20 mice） and experimental group （160 mice）. HSK models were established by scratching on the surface of the cornea followed by inoculation of 1 x 106 plague forming unit（PFU） herpes simplex virus type 1 （HSV-1） KOS strain. Change of ocular surface was examined under the slit lamp biomicroscopy before and 1 day,3,5,7,8,10,14,21 days after inoculation of HSV-1. The corneas of mice were collected on the time points mentioned above. The relative expression level of IL-9 mRNA in the cornea of mice was detected by real-time PCR. Immunocytochemical localization of IL-9 protein in murine cornea was viewed by a confocal microscope after preparation of the corneal cryostat sections. All experimental manipulations were undertaken in accordance with the institutional guidelines for the care and use of laboratory animals. Results Punctiform, dendriform or geographic defect in corneal epithelium were seen 3 days and healing 6 days after inoculation of HSV-1. However, edema and opacity of the corneal stroma appeared 7 days and peaked 14 days following the inoculation. Real-time PCR assay showed that very little of IL-9 mRNA （A26o/A2so ） was expressed in the cornea in the mice of the control group. But in 1 day,3,5,7,8,10 and 14 days, the relative level of IL-9 mRNA was 6.37 ~0. 45,5.66 ＋0.53,3.93 ~0.35,3.62 ~0.34,3.23 ~0. 18,2.57 ~0. 14,2. 19 ~ 0. 20,with a significant difference among the various time points （F = 92. 764,P = 0. 000） , and IL-9 mRNA in 1 day, 3,5,7,8,10 and

关 键 词：I型单纯疱疹病毒 角膜 白细胞介素一9 角膜基质炎 

分 类 号：R77[医药卫生—眼科]