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[摘要]
目的研究地锦草提取物体外抗真菌活性,评价其抗真菌作用,探讨其抗真菌作用机制.方法参照美国临床实验室标准化委员会(NCCLS)推荐的《产孢丝状真菌的液基稀释法抗真菌药物敏感性试验方案》(M38-A),测定地锦草提取物对60株临床常见皮肤癣菌的最小抑菌浓度(MIC)值,扫描电镜和透射电镜下观察其细胞表面形态结构变化和细胞内超微结构变化.结果地锦草提取物对红色毛癣菌的平均MIC为446μg/mL,对石膏样毛癣菌的平均MIC为539μg/mL;扫描电镜下观察地锦草提取物作用于皮肤癣菌后,细胞表面皱缩不平,有严重皱褶、破裂现象;透射电镜下可见,真菌细胞壁不完整,局部有缺损,厚薄不均;细胞膜轮廓不清,局部有破损;胞内细胞器损伤严重,多见空泡化,细胞内成分聚集成电子密度较高的团块.结论地锦草提取物对真菌的生长具有显著地抑制作用,可使其形态和超微结构发生明显的改变.
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[Abstract]
Objective By measuring the minimal inhibitory concentration(MIC)ofEuphorbia humifusaextractin vitroto examine its anti-fungal effect and observe the effect ofE.humifusaextract on the ultrastructure of dermatophytes,so as to explore the potential mechan-ism of its antifungal action.Methods Appling the standard approved by the National Committee for Clinical Laboratory Standards(NCCLS): Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi(M38-A)to investigating the MICs ofE.hum-ifusa extract against 60 strains of common clinical dermatophytes.Treated the sample with the routine method,the ultrastructure of derma-tophytes was observed with scanning electron microscope(SEM)and transmission electron microscope(TEM).Results The mean MIC ofE.hum-ifusa extract againstTrichophyton rubrumwas 446 μg/mL;The mean MIC ofE.humifusaextract againstTrichophyton mentagrophy-teswas 539 μg/mL.After exposing toE.humifusaextract,cell surface wrinkled,and folds and cell breakage were observed under SEM.The cell wall was incomplete,the thickness of cell wall became uneven,the density of the cell wall decreased,and the cell membrane became disco-ntinuous with TEM observation.The organelles such as mitochondria and endoplasmic reticulum were deeply damaged.The cytoplasma was aggregated and some high density electron light areas as well as vacuoles were formed.ConclusionE.humifusaextract has a strong antifungal effect which could obviously change the conformation and the ultrastructure of dermatophytes.
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[基金项目]
国家自然科学基金资助项目(30760308)