摘要
目的为了获得体外抑菌活性较好的抗真菌药物,筛选了14种唑类化学合成药物(A1~A7和B1~B7)的体外抗真菌活性。以白色念珠菌为代表试验菌株,初步探讨其抗真菌机制。方法通过微量液体稀释法测定药物的最小抑菌浓度(minimum inhibitory concentration,MIC),最小杀菌浓度(minimum fungicidal concentration,MFC),琼脂平板计数法测定时间杀菌曲线;棋盘稀释法测定联合用药,评估化合物体外抑菌效果,筛选获得高效抗真菌化合物;以化合物B7为基础,通过液体菌丝诱导观察白色念珠菌菌体形态变化,通过XTT法测定生物被膜形成抑制,通过真菌细胞内氧化应激活性氧初级荧光测定,初步探究化合物对白色念珠菌的体外抗真菌活性及其作用机制。结果本研究从14种唑类化合物中筛选获得具有良好体外抗真菌效果化合物B7,以白色念珠菌ATCC SC5314为代表,其MIC为0.0625μg·mL^(-1),MFC为0.25μg·mL^(-1),且其杀菌活性具有剂量依赖性的特点,与氟康唑联合具有累加作用。菌丝诱导实验表明B7对菌体菌丝生长具有明显抑制作用,XTT实验表明,化合物B7对生物被膜具有一定抑制作用。进行真菌细胞内氧化应激活性氧初级荧光测定,发现随浓度增加,化合物B7均能使白色念珠菌氧化应激活性氧升高且总高于咪康唑阳性对照组。结论化合物B7体外抗真菌效果良好,且能够抑制菌丝生长与生物被膜形成。
Objective In order to obtain drugs with good antibacterial activity in vitro,the antifungal activity of 14 kinds of azole chemical synthetic drugs(A1-A7 and B1-B7)was screened.The antifungal mechanism of Candida albicans was discussed.Methods The minimum inhibitory concentration(MIC)and minimum fungicidal concentration(MFC)of the drug were measured by microliquid dilution method.Time sterilization curve was measured by AGAR plate counting method.The checkerboard dilution method was used to determine the antibacterial effect of the compound in vitro,and the highly effective antifungal compound was screened.Based on compound B7,the morphological changes of Candida albicans were observed through the induction of liquid mycelia,the inhibition of biofilm formation was measured by XTT method,and the in vitro antifungal activity of the compound against Candida albicans and its mechanism of action were preliminarily investigated by the primary fluorescence determination of oxidative stress reactive oxygen species in fungal cells.Results In this study,compound B7 was screened from 14 azole compounds with good antifungal effects in vitro,represented by Candida albicans ATCC SC5314,whose MIC was 0.0625μg·mL^(-1),and MFC was 0.25μg·mL^(-1),and its bactericidal activity was dose-dependent.The combination with fluconazole has additive effect.The mycelium induction experiment showed that B7 had obvious inhibitory effect on mycelium growth,and the XTT experiment showed that compound B7 had certain inhibitory effect on biofilm.The primary fluorescence determination of oxidative stress reactive oxygen species in fungal cells showed that compound B7 could increase the oxidative stress reactive oxygen species of Candida albicans with increasing concentration and was always higher than that of miconazole positive control group.Conclusion Compound B7 has good antifungal effect in vitro,and can inhibit mycelium growth and biofilm formation.
作者
索菲亚珊
牟睿瑶
单路瑶
蔡依依
李辰昊
张雨晴
苏昕
SUO Feiyashan;MOU Ruiyao;SHAN Luyao;CAI Yiyi;LI Chenhao;ZHANG Yuqing;SU Xin(School of Life Sciences and Biopharmaceuticals,Shenyang Pharmaceutical University,Shenyang 110016,China)
出处
《沈阳药科大学学报》
CAS
CSCD
2024年第4期457-463,共7页
Journal of Shenyang Pharmaceutical University
基金
辽宁省科技厅民生科技计划资助项目(2021JH2/10300055)。
关键词
白色念珠菌
致病机制
体外抗菌
candida albicans
pathogenic mechanism
antibacterial in vitro
