PDF(Pubmed)
Abstract：
The antifungal activity of fisetin against Candida albicans is explored, elucidating a mechanism centered on membrane permeabilization and ensuing disruption of pH homeostasis. The Minimum Inhibitory Concentration (MIC) of fisetin, indicative of its interaction with the fungal membrane, increases in the presence of ergosterol. Hoechst 33342 and propidium-iodide staining reveal substantial propidium-iodide accumulation in fisetin-treated C. albicans cells at their MIC, with crystal violet uptake assays confirming fisetin-induced membrane permeabilization. Leakage analysis demonstrates a significant release of DNA and proteins in fisetin-treated cells compared to controls, underscoring the antifungal effect through membrane disruption. Green fluorescence, evident in both the cytoplasm and vacuoles of fisetin-treated cells under BCECF, AM staining, stands in contrast to controls where only acidic vacuoles exhibit staining. Ratiometric pH measurements using BCECF, AM reveal a noteworthy reduction in intracellular pH in fisetin-treated cells, emphasizing its impact on pH homeostasis. DiBAC4(3) uptake assays demonstrate membrane hyperpolarization in fisetin-treated cells, suggesting potential disruptions in ion flux and cellular homeostasis. These results provide comprehensive insights into the antifungal mechanisms of fisetin, positioning it as a promising therapeutic agent against Candida infections.
摘要：
探索了非瑟素对白色念珠菌的抗真菌活性，阐明以膜透化和随后的pH稳态破坏为中心的机制。非瑟酮的最低抑菌浓度(MIC),表明它与真菌膜的相互作用，麦角甾醇的存在增加。Hoechst33342和碘化丙啶染色显示，在非瑟酮处理的白色念珠菌细胞的MIC下，碘化丙啶大量积累，结晶紫摄取测定证实了非瑟酮诱导的膜透化。泄漏分析表明，与对照组相比，非瑟酮处理的细胞中DNA和蛋白质的显着释放。强调通过膜破坏的抗真菌作用。绿色荧光,在BCECF下，Fisetin处理的细胞的细胞质和液泡中都很明显，AM染色,与只有酸性液泡表现出染色的对照相反。使用BCECF进行比例pH测量，AM显示非塞素处理细胞的细胞内pH值显著降低，强调其对pH稳态的影响。DiBAC4（3）摄取测定证明非瑟酮处理的细胞中膜超极化，提示离子流和细胞稳态的潜在破坏。这些结果为非塞素的抗真菌机制提供了全面的见解，将其定位为针对念珠菌感染的有前途的治疗剂。
