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【目的】研究在体外情况下和厚朴酚对白色念珠菌的抑制作用及其可能机制。【方法】采用微量稀释法测定和厚朴酚对白色念珠菌的最低抑菌浓度(MIC80)和最低杀菌浓度(MFC)用透射电镜观察不同浓度和厚朴酚对白色念珠菌超微结构的影响;采用AnnexinⅤ-FITC/PI染色法分析不同浓度和厚朴酚对白色念珠菌细胞凋亡的影响;用DCFH-DA染色法测定不同浓度和厚朴酚对白色念珠菌细胞内活性氧积累的影响;用JC-1染色法分析不同浓度和厚朴酚对白色念珠菌线粒体膜电位的影响;用碘化丙啶染色、考马斯亮蓝G-250染色检测和厚朴酚对白色念珠菌细胞膜通透性的影响;通过测定加入麦角甾醇后,和厚朴酚对白色念珠菌的抑制作用的变化,检测和厚朴酚对白色念珠菌细胞膜的影响;【结果】和厚朴酚对白色念珠菌具有很强的抑制作用,MIC和MFC分别为16μg/mL和32μg/mL。对白色念珠菌细胞壁、细胞膜和胞浆均有明显的影响。和厚朴酚是通过增加活性氧的产生和破坏线粒体功能来诱导白念珠菌的细胞凋亡和坏死。它也影响细胞膜的通透性,这可能和细胞壁的破坏和与麦角固醇的结合有关。【结论】结果表明,和厚朴酚通过产生活性氧并伴随着一系列的细胞损伤这种复杂的机制从而对白色念珠菌产生抑制作用,使和厚朴酚成为一种潜在的抗真菌药物。
【Objective】 To investigate the inhibitory effect of honokiol on Candida albicans in vitro and its possible mechanism. 【Method】 The minimum inhibitory concentration (MIC80) and minimum bactericidal concentration (MFC) of honokiol on Candida albicans were determined by microdilution method. The effects of different concentrations of honokiol on the ultrastructure of Candida albicans were observed by transmission electron microscopy ; Annexin Ⅴ-FITC / PI staining analysis of different concentrations of honokiol and Candida albicans cell apoptosis; using DCFH-DA staining determination of different concentrations of honokiol and Candida albicans intracellular reactive oxygen species accumulation The effect of different concentrations of honokiol on the mitochondrial membrane potential of Candida albicans was analyzed by JC-1 staining. The membrane permeability of Candida albicans was detected by propidium iodide staining and Coomassie brilliant blue G-250 staining The effect of honokiol on Candida albicans cell membrane was tested by measuring the inhibitory effect of honokiol on Candida albicans after addition of ergosterol. 【Results】 Honokiol has the inhibitory effect on Candida albicans Strong inhibition, MIC and MFC were 16μg / mL and 32μg / mL. Candida albicans cell wall, cell membrane and cytoplasm have a significant impact. Honokiol induces C. albicans apoptosis and necrosis by increasing the production of reactive oxygen species and destroying mitochondrial function. It also affects the permeability of the cell membrane, which may be related to cell wall destruction and ergosterol binding. [Conclusion] The results show that honokiol is a potential antifungal agent that inhibits Candida albicans by generating a complex mechanism of reactive oxygen species with a series of cell damage.