兔曲霉菌性角膜炎動物模型建立及角膜共焦顯微鏡檢查

1、兔曲霉菌性角膜炎動物模型建立及角膜共焦顯微鏡檢查            兔曲霉菌性角膜炎動物模型建立及角膜共焦顯微鏡檢查    2008-5-9 12:13:29                       &

2、#160;                                       作者：杜蕊,吳潔,馬吉獻,朱秀萍,劉先寧,尹琳,程燕    【關(guān)鍵詞】  真菌性角膜炎；曲霉

3、菌；兔;動物模型；角膜共焦顯微鏡    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：建立有色實驗兔真菌性角膜炎動物模型，角膜

9、共焦顯微鏡進行早期診斷. 方法：應(yīng)用內(nèi)皮面作“#”形劃痕的角膜植片，用100尼龍縫線將其固定于受體角膜上，將曲霉菌混懸液注入植片與植床間，建立模型. 并在接種后第3，7，10，15日作裂隙燈、角膜刮片、角膜共焦顯微鏡、角膜組織病理切片檢查及培養(yǎng). 結(jié)果：兔眼真菌性角膜炎動物模型成功建立，并經(jīng)角膜刮片、角膜共焦顯微鏡、組織病理切片染色檢查及培養(yǎng)證實. 共焦顯微鏡檢查早期（接種3 d）見較直、短、分支比較少的高反光的菌絲及孢子，菌絲以斜行、垂直生長方式為主，伴有少量的炎癥細(xì)胞浸潤. 中期（接種7,10 d）共焦顯微鏡檢查除可見高反光的菌絲外還可見大量的炎癥細(xì)胞（中性粒細(xì)胞、巨噬細(xì)胞）浸潤. 后期（

10、接種15 d）在角膜刮片和組織病理切片檢查均未見明顯真菌菌絲時，共焦顯微鏡檢查仍可在基質(zhì)深層見到少量稀疏分布、短小的真菌菌絲. 結(jié)論：應(yīng)用角膜植片成功建立兔曲霉菌性角膜炎模型，共焦顯微鏡為曲霉菌性角膜炎早期診斷提供了重要的客觀依據(jù).        【關(guān)鍵詞】 真菌性角膜炎；曲霉菌；兔;動物模型；角膜共焦顯微鏡         0引言        真菌性角膜炎(fungal keratitis, FK)或稱角膜真菌病(keratomycosis)致盲率極高，尤其在西北地區(qū). 近幾年來，由于廣譜抗生素、糖皮質(zhì)激素和免疫抑制劑的廣泛使用及角膜接觸鏡佩戴人群的增多，其發(fā)病率明顯升高1-4，在某些地區(qū)甚至已躍居感染性角膜疾病的首位5-6. 查閱近20年的文獻，未發(fā)現(xiàn)有通過角膜劃痕等方法成功建立真菌性角膜炎模型的報道. 傳統(tǒng)的層間注入法和軟性接觸鏡法與真菌性角膜炎的臨床病程不同. 本實驗我們不使用糖皮質(zhì)激素，應(yīng)用角膜植片，建立了一種兔真菌性角膜炎的模型，并應(yīng)用角膜共焦顯微鏡