茄病镰刀菌性角膜炎树鼩模型的建立  

Establishment of a tree shrew model of Fusarium keratitis

作　　者：李妍[1] 代解杰[2] 孙晓梅[2] 刘海[1] 胡竹林[1] 

机构地区：[1]昆明医科大学第四附属医院眼科,云南省第二人民医院眼科,云南省眼科研究所,云南省眼科疾病防治研究重点实验室,昆明650021 [2]中国医学科学院/北京协和医学院医学生物学研究所,昆明650118

出　　处：《中国实验动物学报》2017年第4期420-424,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：云南省卫生科技计划项目(2014NS040,2017NS129);昆明医科大学博士创新基金(2016D07)

摘　　要：目的采用临床分离的茄病镰刀菌感染树鼩角膜,建立茄病镰刀菌性角膜炎树鼩模型。方法茄病镰刀菌接种到沙保氏培养基,26℃培养箱培养7 d,收集真菌混悬液,血细胞计数板调整孢子数量为1×10^(10)CFU/m L。清洁级树鼩40只随机分为实验组(n=30)、对照组(n=10)。实验组用胰岛素针头(29 G)将真菌孢子混悬液50μL注入角膜基中央,对照组注入生理盐水50μL。通过前段照相、共聚焦显微镜检查、病理组织学变化、感染角膜组织培养对模型进行评价。结果真菌浸润范围、角膜上皮细胞和内皮细胞水肿程度、菌丝数量均与时间呈正相关;炎性细胞浸润数量造模后第7天达到高峰,以中性粒细胞为主;实验各时间点均可见菌丝平行于基质纤维生长;感染后角膜组织培养可见茄病镰刀菌生长;造模成功率为86%。结论采用基质注射茄病镰刀菌孢子的方法首次成功建立茄病镰刀菌性角膜炎树鼩模型。Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma. Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days. Fungal suspension was collected and the number of spores was adjusted to 1 × 1010CFU/m L on the blood cell count plate. Forty healthy tree shrews were randomly divided into experimental group（ n = 30） and control group（ n = 10）. In the experimental group,50 μL of fungal spore suspension was injected into the cornea center with a 29 G needle,and 50 μL saline was injected in the control group. The models were evaluated by anterior segment photography,in vivo confocal microscopy,histopathology,and corneal tissue culture. Results The fungal infiltration,the degree of edema of corneal epithelial and endothelial cells,and the number of mycelium were positively correlated with time. The number of infiltrating inflammatory cells,mainly,neutrophils,reached a peak on the 7th day after modeling. The mycelial growth was parallel to the stromal fibers. After the successful establishment of the model,the corneal tissue culture showed the growth of Fusarium solani. The successful rate of modeling was 86%. Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.

关 键 词：树鼩 茄病镰刀菌 动物模型 真菌感染 角膜炎 

分 类 号：Q95-33[生物学—动物学]