Jilin Provincial Key Laboratory on Molecular and Chemical Genetics, The Second Hospital of Jilin University, Changchun 130024, China.
Eye Center, The Second Hospital of Jilin University, Changchun 130024, China.
J Mycol Med. 2018 Mar;28(1):36-44. doi: 10.1016/j.mycmed.2017.12.011. Epub 2018 Feb 22.
The aim of this study was to investigate the antifungal activity of dracorhodin perchlorate (DP) against planktonic growth and virulence factors of Candida albicans.
Microdilution method based on CLSI-M27-A3 was used to test the antifungal susceptibility of DP. The activity of DP against biofilm formation and development of C. albicans was quantified by XTT assay and visualized by confocal laser scanning microscope. The effect of DP on the morphological transition of C. albicans induced by four kinds of hyphal-inducing media at 37°C for 4hours was observed under microscope. The rescue experiment by adding exogenous cAMP analog was performed to investigate the involvement of cAMP in the yeast to hyphal transition and biofilm formation of C. albicans. Egg yolk emulsion agar was used to determine the inhibition of DP on the phospholipase production of C. albicans. Human JEG-3 and HUVEC cell lines, as well as the nematode Caenorhabditis elegans was used to assess the toxicity of DP.
The minimum inhibitory concentration (MIC) of DP is 64μM while the antifungal activity was fungistatic. As low as a concentration at 16μM, DP could inhibit the yeast to hyphal transition in liquid RPMI-1640, Spider, GlcNAc and 10% FBS-containing Sabouroud Dextrose medium, as well as on the solid spider agar. Exogenous cAMP analog could rescue part of biofilm viability of C. albicans. DP could inhibit the production of phospholipase. The toxicity of DP against human cells and C. elegans is low.
DP could inhibit the planktonic growth and virulent factors in multiple stages, such as yeast to hyphal transition, adhesion, biofilm formation and production of phospholipase of C. albicans.
本研究旨在探讨血根碱高氯酸盐(DP)对白色念珠菌浮游生长和毒力因子的抗真菌活性。
采用基于 CLSI-M27-A3 的微量稀释法检测 DP 的抗真菌药敏性。通过 XTT 测定和共聚焦激光扫描显微镜观察来量化 DP 对白色念珠菌生物膜形成和发育的抑制作用。在显微镜下观察 DP 对四种诱导菌丝形成的培养基在 37°C 下 4 小时诱导的白色念珠菌形态转变的影响。通过添加外源性 cAMP 类似物进行挽救实验,以研究 cAMP 在白色念珠菌酵母到菌丝过渡和生物膜形成中的作用。使用卵黄乳液琼脂测定 DP 对白色念珠菌磷脂酶产生的抑制作用。用人 JEG-3 和 HUVEC 细胞系以及线虫秀丽隐杆线虫来评估 DP 的毒性。
DP 的最低抑菌浓度(MIC)为 64μM,而抗真菌活性为抑菌作用。DP 浓度低至 16μM 时,即可抑制 RPMI-1640 液体、蜘蛛、GlcNAc 和 10% FBS 含 Sabouroud Dextrose 培养基中的酵母到菌丝过渡,以及固体蜘蛛琼脂中的酵母到菌丝过渡。外源性 cAMP 类似物可以部分挽救白色念珠菌生物膜活力。DP 可以抑制磷脂酶的产生。DP 对人细胞和秀丽隐杆线虫的毒性较低。
DP 可以抑制白色念珠菌的浮游生长和多个阶段的毒力因子,如酵母到菌丝过渡、黏附、生物膜形成和磷脂酶产生。