兔曲霉菌性角膜炎动物模型建立及角膜共焦显微镜检查

1、兔曲霉菌性角膜炎动物模型建立及角膜共焦显微镜检查            兔曲霉菌性角膜炎动物模型建立及角膜共焦显微镜检查    2008-5-9 12:13:29                       &

2、#160;                                       作者：杜蕊,吴洁,马吉献,朱秀萍,刘先宁,尹琳,程燕    【关键词】  真菌性角膜炎；曲霉

3、菌；兔;动物模型；角膜共焦显微镜    Establishment of rabbit fungal (aspergillus) keratitis model and corneal examination with confocal microscope        【Abstract】 AIM: To establish a rabbit model of fungal (aspergillus) keratitis and evaluate the role of confo

4、cal microscopy in early diagnosis. METHODS: Cornea slice, with a “#”shaped nick on its endothelium, was sewed onto acceptor cornea with 100 nylon thread. Aspergillus suspensions was injected into the space between cornea slice and acceptor cornea. Slit lamp microscopy, cornea scraping, confocal micr

5、oscope, pathological section examination and culture were used at day 3, 7, 10 and 15  after aspergillus was inoculated. RESULTS: A rabbit model of fungal keratitis was built successfully and proved by cornea scraping, confocal microscopy, pathological section examination and culture.  In

6、the early period (3 d after aspergillus injection), the straight, short, fewbranches and dense lightreflecting hypha, spore, and a little inflammatory infiltration  were found by confocal microscopy. In the metaphase (7, 10 d after aspergillus injection), a lot of hypha and inflammatory infiltr

7、ation were discovered by confocal microscopy. In the anaphase (15 d  after aspergillus injection), a little hypha was found by confocal microscopy while it could not be found by cornea scraping and pathological section examination. CONCLUSION: A rabbit model of fungal keratitis was established

8、successfully. Confocal microscopy provides an important basis for early diagnosis of fungal keratitis.        【Keywords】 fungal keratitis; aspergillus; rabbits; animal model; confocal microscope        【摘要】 目的：建立有色实验兔真菌性角膜炎动物模型，角膜

9、共焦显微镜进行早期诊断. 方法：应用内皮面作“#”形划痕的角膜植片，用100尼龙缝线将其固定于受体角膜上，将曲霉菌混悬液注入植片与植床间，建立模型. 并在接种后第3，7，10，15日作裂隙灯、角膜刮片、角膜共焦显微镜、角膜组织病理切片检查及培养. 结果：兔眼真菌性角膜炎动物模型成功建立，并经角膜刮片、角膜共焦显微镜、组织病理切片染色检查及培养证实. 共焦显微镜检查早期（接种3 d）见较直、短、分支比较少的高反光的菌丝及孢子，菌丝以斜行、垂直生长方式为主，伴有少量的炎症细胞浸润. 中期（接种7,10 d）共焦显微镜检查除可见高反光的菌丝外还可见大量的炎症细胞（中性粒细胞、巨噬细胞）浸润. 后期（

10、接种15 d）在角膜刮片和组织病理切片检查均未见明显真菌菌丝时，共焦显微镜检查仍可在基质深层见到少量稀疏分布、短小的真菌菌丝. 结论：应用角膜植片成功建立兔曲霉菌性角膜炎模型，共焦显微镜为曲霉菌性角膜炎早期诊断提供了重要的客观依据.        【关键词】 真菌性角膜炎；曲霉菌；兔;动物模型；角膜共焦显微镜         0引言        真菌性角膜炎(fungal keratitis, FK)或称角膜真菌病(keratomycosis)致盲率极高，尤其在西北地区. 近几年来，由于广谱抗生素、糖皮质激素和免疫抑制剂的广泛使用及角膜接触镜佩戴人群的增多，其发病率明显升高1-4，在某些地区甚至已跃居感染性角膜疾病的首位5-6. 查阅近20年的文献，未发现有通过角膜划痕等方法成功建立真菌性角膜炎模型的报道. 传统的层间注入法和软性接触镜法与真菌性角膜炎的临床病程不同. 本实验我们不使用糖皮质激素，应用角膜植片，建立了一种兔真菌性角膜炎的模型，并应用角膜共焦显微镜