免疫抑制BALB/c小鼠侵袭性白念珠菌感染动物模型的建立  

Establishment of an animal model of invasive Candida albicans infection in immunosuppressive BALB/c mice

作　　者：邓洁华[1] 王刚生[1] 张晓光[1] 罗静娜 王佳[3] 祁晓明 李艳玲[1] DENG Jie-hua;WANG Gang-sheng;ZHANG Xiao-guang;LUO Jing-na;WANG Jia;QI Xiao-ming;LI Yan-ling(Department of Dermatology and Venereology,Second Hospital of Hebei Medical University,Hebei Dermatology and Venereology Clinical Medicine Research Center,National Dermatology and Immune Disease Clinical Medicine Research Center Sub-Center Construction Unit,Shijiazhuang 0500009 China)

机构地区：[1]河北医科大学第二医院皮肤性病科,河北省皮肤性病临床医学研究中心,国家皮肤病与免疫病临床医学研究中心分中心建设单位,石家庄05000 [2]河北医科大学第二医院转化医学中心 [3]河北医科大学第二医院质量控制处 [4]河北医科大学第二医院电镜室

出　　处：《医学动物防制》2021年第9期838-841,F0003,共5页Journal of Medical Pest Control

基　　金：河北省自然科学基金(H2013206316)。

摘　　要：目的建立免疫抑制BALB/c小鼠侵袭性白念珠菌感染的动物模型。为研发新型靶向抗真菌药物提供依据。方法对BALB/c小鼠腹腔注射环磷酰胺200 mg/kg,每日1次,连续2次,第4天经鼻腔灌注白念珠菌菌悬液50μl(107 cfu/ml),第7天取肺组织标本进行真菌直接镜检、培养、病理检查、(1,3)-β-D-葡聚糖检测及电镜观察。结果小鼠被注射环磷酰胺的第4天,白细胞2.48×10^(9)/L、中性粒细胞16.8×10^(9)/L降至最低值。免疫抑制的小鼠经鼻腔灌注白念珠菌菌悬液后的第7天,取肺组织标本进行真菌镜检,镜下可见大量菌丝及芽生孢子,沙堡培养有白念珠菌生长,组织病理可见大量菌丝及芽生孢子及炎性细胞浸润、坏死,(1,3)-β-D-葡聚糖检测(5 181.33±223.67)pg/ml与生理盐水对照组(75.62±18.34)pg/ml比较,差异有统计学意义(t=-4.536,P<0.01),透射电镜可见白念珠菌细胞壁为多层板状结构、清晰的细胞膜及细胞器。结论对免疫抑制BALB/c小鼠鼻腔灌注白念珠菌菌悬液后,经取肺组织标本进行真菌检查、培养、病理检查、(1,3)-β-D-葡聚糖检测及电镜观察方法,成功建立了免疫抑制BALB/c小鼠侵袭性白念珠菌感染动物模型,为观察药物在体内对真菌细胞壁及(1,3)-β-D-葡聚糖的影响提供了依据。Objective To establish an animal model for invasive Candida albicans infection in immunosuppressive BALB/c mice, and to provide the basis for development of new targeted antifungal drugs.Methods BALB/c mice were injected intraperitoneally with 200 mg/kg cyclophosphamide for 2 consecutive times once daily, 50 μl Candida albicans suspension(107 cfu/ml) was injected via nasal drip on the fourth day, lung tissue specimens were implemented on the seventh day, and direct microscopic examination, culture, pathological examination,(1,3)-β-D-glucan detection and electron microscopic observation of fungi were implemented.Results The white blood cells in 2.48×10^(9)/L and the neutrophils in 16.8×10^(9)/L were decreased to the lowes on the 4 th day after the mice were injected with cyclophosphamide, and the lung tissue specimens were taken for fungal microscopic examination on the 7 th day after the immunosuppressed mice were perfused with Candida albicans suspension through nasal cavities.Abundant hyphae and spores were found under the microscope, Candida albicans grew and were cultured in the sandcastle, massive hyphae, spores, inflammatory cell infiltration and necrosis in histological-pathological region, and the difference between(1,3)-β-D-glucan detection(5 181.33±223.67)pg/ml and the normal saline control group(75.62±18.34)pg/ml was statistically significant(t=-4.536,P<0.01),transmission electron microscope observation showed that the cell wall of Candida albicans was featured with multilayer plate structure, transparent cell membrane and organelles.Conclusion Lung tissue samples are taken for fungal examination, culture, pathological examination,(1,3)-β-D-glucan detection and electron microscopy observation method when Candida albicans suspension are perfused via nasal cavities of immunosuppressive BALB/c mice, successfully setting up an animal model of invasive Candida albicans infection in immunosuppressive BALB/c mice, and providing the basis for observing the effects of drugs on fungal cell walls and(1,3)

关 键 词：侵袭性念珠菌感染 (1 3)-β-D-葡聚糖 真菌镜检和培养 病理检查 透射电镜 动物模型 

分 类 号：R379.4[医药卫生—病原生物学]