CAJ | 학술논문

目的构建无绿藻感染小鼠皮肤模型.方法 BALB/c小鼠分为4组,高浓度接种免疫抑制组为免疫抑制小鼠,腹部皮下接种1×109孢子/ml中型无绿藻波多黎各变种悬液组；低浓度接种免疫抑制组为免疫抑制小鼠,腹部皮下接种1×106孢子/ml悬液组；高浓度接种健康组为健康小鼠,腹部皮下接种1×109孢子/ml悬液组；对照组为健康小鼠腹部皮下接种生理盐水溶液,每个部位接种量均为200μl.接种后第7、14、28天时观察各组小鼠腹部皮损变化,并进行腹部皮肤病理及真菌检查.结果实验组小鼠接种处均出现丘疹、脓肿,部分出现溃疡、结痂,感染率均为100％.3组小鼠的腹部皮损直径,组内各时间点比较差异均有统计学意义(均P＜ 0.05),均在第7天时最大,分别为(6.75±1.09) mm、(5.88±1.17) mm和(5.96±0.99)mm;组间相比,在接种后第7、14天时差异无统计学意义(P＞0.05),在第28天时差异有统计学意义(F=8.91,P＜0.05),高浓度接种免疫抑制组最大为(4.38±0.86) mm.3个实验组基本病理改变为坏死、脓肿、肉芽肿形成,HE染色和过碘酸雪夫(PAS)染色均可见孢子,皮损直接镜检可见孢子,培养为无绿藻生长.对照组未感染无绿藻.结论腹部皮下接种免疫抑制和健康小鼠均可构建无绿藻病皮肤感染模型.
목적 구건무록조감염소서피부모형.방법 BALB/c소서분위4조,고농도접충면역억제조위면역억제소서,복부피하접충1×109포자/ml중형무록조파다려각변충현액조；저농도접충면역억제조위면역억제소서,복부피하접충1×106포자/ml현액조；고농도접충건강조위건강소서,복부피하접충1×109포자/ml현액조；대조조위건강소서복부피하접충생리염수용액,매개부위접충량균위200μl.접충후제7、14、28천시관찰각조소서복부피손변화,병진행복부피부병리급진균검사.결과 실험조소서접충처균출현구진、농종,부분출현궤양、결가,감염솔균위100％.3조소서적복부피손직경,조내각시간점비교차이균유통계학의의(균P＜ 0.05),균재제7천시최대,분별위(6.75±1.09) mm、(5.88±1.17) mm화(5.96±0.99)mm;조간상비,재접충후제7、14천시차이무통계학의의(P＞0.05),재제28천시차이유통계학의의(F=8.91,P＜0.05),고농도접충면역억제조최대위(4.38±0.86) mm.3개실험조기본병리개변위배사、농종、육아종형성,HE염색화과전산설부(PAS)염색균가견포자,피손직접경검가견포자,배양위무록조생장.대조조미감염무록조.결론 복부피하접충면역억제화건강소서균가구건무록조병피부감염모형.
Objective To develop a mouse model of cutaneous protothecosis.Methods Totally,48 BALB/c mice were randomly and equally divided into four groups:high-and low-concentration immunosuppressive groups-immunosuppressed mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 and 1 × 106 colony forming units (CFU) conidia/ml respectively,high-concentration healthy group-healthy mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 CFU conidia/ml,and control group-healthy mice inoculatedwith sodium chloride physiological solution.The P.zopfii suspension or sodium chloride physiological solution was subcutaneously inoculated to the abdominal skin of mice,with the inoculation volume being 200 μl.Skin appearance at the inoculation site was observed,and four mice were sacrificed in each group on day 7,14 and 28 after the inoculation.Skin specimens were resected from the inoculation sites of mice and subjected to pathological and mycological examinations.Results Mice in the three experiment groups inoculated with the P.zopfi suspension were all infected,with the appearance of papules and abscess at the inoculation sites of all mice as well as ulcer and crusts in some mice.Meanwhile,no mice were infected in the control group.Significant differences were noted in the diameter of skin lesions between the three time points in these experiment groups (all P ＜ 0.05),and the largest diameter of lesions was observed on day 7,which was (6.75 ± 1.09) mm in the high-concentration immunosuppressive group,(5.88 ± 1.17) mm in the low-concentration immunosuppressive group,and (5.96 ± 0.99) mm in the high-concentration healthy group.Comparisons of lesion diameter between the three experiment groups revealed a statistical difference on day 28 (F =8.91,P ＜ 0.05),with the largest lesion diameter observed in the high-concentration immunosuppressive group (4.38 ± 0.86 mm),but no statistical difference was found on day 7 or 14 (both P ＞ 0.05).Pathology of skin specimens consistently revealed necrosis,abscess and granuloma formation in the three experiment groups.Spores were found in the lesions of infected mice by haematoxylin-eosin staining,periodic acid-Schiff staining,and direct microscopy.Culture of tissue samples from the experiment groups grew Prototheca.Conclusion The mouse model of protothecosis can be established by subcutaneous inoculation of Prototheca suspension in the abdominal skin of healthy or immunosuppressed mice.