小鼠真菌性角膜炎中主要免疫细胞的作用  

Effects of immunocyte on the process of fungal keratitis

作　　者：张红敏[1] 刘素素[1] 许中中[1] 马聪慧[1] 谢艳亭[1] 吴细丕[1] 王丽娅[1] 

机构地区：[1]河南省眼科研究所 河南省角膜病重点实验室,郑州450003

出　　处：《中华实验眼科杂志》2012年第9期779-784,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81170831）;河南省基础与前沿计划项目（102300410024、112300410036、11230040093）

摘　　要：背景在中国，感染性角膜病变是引起角膜盲的主要原因之一，而其中真菌性角膜炎的发病率高、危害大，是角膜盲防治工作的重点。研究表明，真菌性角膜炎后局部的免疫状态在疾病的发生发展过程中起着至关重要的作用，但其免疫机制尚待进一步研究。目的研究真菌性角膜炎发生发展过程中局部免疫细胞的作用及其机制。方法共选用SPF级12周龄雄性C57BL／6J小鼠48只，按照随机数字表法将小鼠随机分为对照组和模型组，每组24只小鼠，然后根据检测的时间点将成模的小鼠随机亚分为临床评分组，6、9、12、24、72和120h组，每组3只小鼠。模型组小鼠用角膜划痕和带有真菌的竹签涂抹法接种腐皮镰孢菌以建立真菌性角膜炎模型，对照组仅用无菌刀片划痕而不接种任何真菌。各组小鼠分别于上述时间点进行裂隙灯显微镜检查，采用改良标准对炎症程度进行临床评分。其余组按照时间点处死小鼠，采用免疫荧光标记法对炎症局部中性粒细胞和T淋巴细胞进行鉴定，计数炎性细胞数量。实验动物的使用遵循ARVO声明，研究过程经郑州大学实验动物伦理委员会和河南省眼科研究所生命科学实验管理与伦理审查委员会批准。结果造模成功后24h模型组小鼠出现角膜混浊，72h时病变最为严重，此后病变逐渐减轻，而对照组24h后上皮愈合。造模后24～72h，模型组眼部炎症反应评分逐渐达峰值，与造模后6h相比差异均有统计学意义（P〈0．01）；造模后6、9、12h模型组与对照组比较，角膜炎症反应评分均明显升高，差异均有统计学意义（P〈0．05）。造模后9h和12h模型组小鼠中性粒细胞角膜浸润数量增加，与对照组相比差异均有统计学意义（P〈0．05），与24、72、120h比较差异亦有统计学意义（P=0．000）。模型组造模后12、24、72h角膜中性粒细胞与6h比较差异均有Background Infective keratopathy is a key cause of corneal blindness in China, and fungal keratitis is proved to have a higher incidence and bigger threats in infective keratitis. Researches showed that topical immunology plays an important effect during the development of fungal keratitis ,but its mechanism is still studying. Objective This experiment was to explore the critical immunoeyte during the process of fungal keratitis. Methods Forty-eight SPF 12-week-old male C57BL/6J mice were included and randomized into the control group and model group. The fungal keratitis model closely mimicking human cornea infections was established in the mouse using scratch followed by incubation of fusarium solani on the cornea, and the mice in the control group scratched on the cornea only. Cornea was examined under the slit lamp at 0,6,9,12,24,72 and 120 hours after operation. The severity of keratomycosis was clinically scored based on the literature criteria. The inflammatory cells were identified using immnofluorescence label, and the number of the inflammatory ceils was calculated and compared among different groups and time points. This study complied with the Statement of ARVO in the use of experimental animal. Both Experimental Animal Ethic Commission in Zhengzhou University and Life Science Management Commission approved this study proposal. Results After inoculation of fusarium solani, typical fungul keratitis signs were seen on the cornea. Severe corneal opacification occurred within 24 hours and peaked at 72 hours. However,only mild edema of cornea was exhibited and gradually recovered normal in the control group within 24 hours. The clinical score of inflammation was higher in the model group in various time points than that in the control group, and it was seen that 24-72 hours after operation,the score attached peak in the model group with a significant difference in comparison with the control group（P〈0. 01 ）. In 9,12,24,72 and 120 hours after operation, the number of neutrophil cells was significantly

关 键 词：角膜 真菌/感染 免疫/局部 中性粒细胞 T淋巴细胞 

分 类 号：R772.2[医药卫生—眼科]