丁香酚体外抗阿萨希毛孢子菌的电镜观察

Observationof Eugenol in Vitro Anti-Trichosporon Asahii Electron Microscope

目的观察中药单体——丁香酚(Eugenol)对阿萨希毛孢子菌(Trichosporon asahii,T.asahii)超微结构的影响,探讨其可能存在的潜在作用机制。方法参照NCCLS推荐的M27-A3方案,将受试药物与阿萨希毛孢子菌在35℃下共同培养5d后分离菌体,按常规方法进行样品处理后,扫描电镜和透射电镜下观察其细胞表面形态与细胞内超微结构的变化情况。结果扫描电镜下:丁香酚作用阿萨希毛孢子菌后,真菌的菌体出现肿胀膨大和干瘪变形,部分菌丝的胞壁破坏,随着丁香酚药液浓度的增加,菌丝完全被破坏,且胞质外溢。透射电镜下:丁香酚作用阿萨希毛孢子菌后,真菌的细胞壁不完整,局部有缺损,厚薄不均匀;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。结论丁香酚作用阿萨希毛孢子菌后,可使其形态和超微结构发生明显的改变,证实丁香酚具有抗阿萨希毛孢子菌作用,为开发中药治疗阿萨希毛孢子菌病提供理论依据。

Objective To observe the effects of eugenol on the ultrastructure of Trichosporon asahii （ T. asahii） and to find out the potential mechanism of their antifungal action. Methods According to the National Committee for Clinical Laboratory Standards （NCCLS） M27-A3, we incubated the mixture of eugenol and T. asahii 5 days in foster box of 35~C temperature, and treated the sample under the routine method, finally observed the cell surface morphology and ultrastructure changes of intracellular of T. asahii under scanning electron micro- scope （SEM） as well as transmission electron microscope（TEM）. Results Observed under SEM, after expo- sing to eugenol, cell swelling swollen, shriveled deformation,while part of the mycelium cell wall were dam- aged. Along with the increase of eugenol, we could find that the mycelium were completely destroyed and cy- toplasm were spillover, meanwhile the cell wail became incomplete,the thickness of ceil wall became uneven and the density of the cell wall were decreased. Observed under TEM, after exposing to eugenol, fungal cell wall became incomplete, local defected, and uneven thickness; while membrane contour became unclear, partly damaged;the most serious damage were that common vacuoles appeared in intracellular organelles. Conclusion Eugenol has a strong antifungal effect which can obviously change the conformation and the ul- trastructure of T. asahii. The study confirmed that Eugenol have a great role in anti-T, asahii spores ,and pro- vide a theoretical basis for the development of traditional Chinese medicine treatment of T. asahii.