兔曲霉菌性角膜炎动物模型建立及角膜共焦显微镜检查

Establishment of rabbit fungal (aspergillus) keratitis model and corneal examination with confocal microscope

目的:建立有色实验兔真菌性角膜炎动物模型,角膜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10-0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3d)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7,10d)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(接种15d)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.

AIM ： To establish a rabbit model of fungal （ aspergillus） keratitis and evaluate the role of confocal microscopy in early diagnosis. METHODS ： Cornea slice, with a ＂#＂ -shaped nick on its endothelium, was sewed onto acceptor cornea with 10- 0 nylon thread. Aspergillus suspensions was injected into the space between cornea slice and aoceptor cornea. Slit lamp microscopy, cornea scraping, confocal microscope, 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 the early period （3 d after aspergillus injection ）, the straight, short, few-branches and dense light-reflecting hypha, spore, and a little inflammatory infiltration were found by confocal microscopy. In the metaphase （7, 10 d after aspergillus injecion）, a lot of hypha and inflammatory infiltration 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 successfully, Confocal microscopy provides an important basis for early diagnosis of fungal keratitis.