基于虚拟筛选技术对氟康唑细胞毒性的机制评价  

Mechanism evaluation of fluconazole cytotoxicity based on virtual screening technology

作　　者：李松哲 孙悦 蔡凌云 孙阳[1] LI Songzhe;SUN Yue;CAI Lingyun;SUN Yang(College of Basic Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学基础医学院,哈尔滨150040

出　　处：《中国真菌学杂志》2023年第5期391-396,共6页Chinese Journal of Mycology

基　　金：国家自然科学基金青年基金(81704054);黑龙江省博士后科研启动金(LBH-Q19184)。

摘　　要：目的探讨氟康唑对细胞是否具有毒性,能否用于防治细胞污染。方法虚拟筛选与分子对接技术筛选潜在的毒性靶点。使用人肝癌SMMC-7721细胞、人胚肝HHL-5细胞与人胚肾HEK-293细胞进行体外实验。CCK8实验验证氟康唑对细胞活性的影响。Hoechst 33342/PI双染实验进行形态学观察。qRT-PCR实验进一步验证氟康唑对虚拟筛选所获的靶点是否有影响,明确对细胞影响的机制。结果虚拟筛选前6个核心毒性靶点是IL6、TP53、TNF、IL1β、VEGFA以及EGFR。CCK8实验与Hoechst 33342/PI双染实验显示氟康唑对SMMC-7721细胞和HHL-5细胞具有抑制作用,对HEK-293细胞具有促增殖作用。qRT-PCR实验表明氟康唑导致细胞毒性的核心靶点是IL6、IL1β和EGFR。结论氟康唑不适合广泛应用于预防细胞污染。Objective To investigate whether fluconazole is toxic to cells and whether it can be used to prevent and control cell contamination.Methods Virtual screening and molecular docking technology were used to screen potential toxic targets.In vitro experiments using human hepatoma SMMC-7721 cells,human embryonic liver HHL-5 cells and human embryonic kidney HEK-293 cells were performed.The effect of fluconazole on cell viability was verified by CCK8 assay.Hoechst 33342/PI double staining experiment was used for morphological observation.qRT-PCR experiments further verified whether fluconazole had an effect on the targets obtained by virtual screening,and clarified the mechanism of the effect on cells.Results The first six core toxicity targets of virtual screening were IL6,TP53,TNF,IL1β,VEGFA and EGFR.CCK8 assay and Hoechst 33342/PI double staining assay showed that fluconazole had inhibitory effects on SMMC-7721 cells and HHL-5 cells,and had a proliferative effect on HEK-293 cells.qRT-PCR experiments showed that the core targets of fluconazole-induced cytotoxicity were IL6,IL1βand EGFR.Conclusions Fluconazole is not suitable for widespread use in the prevention of cellular contamination.

关 键 词：氟康唑 虚拟筛选 分子对接 细胞污染 

分 类 号：R978.5[医药卫生—药品]