Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee (IIT-Roorkee), Roorkee, 247667, Uttarakhand, India.
Department of Biotechnology, Graphic Era (Deemed To Be University), Dehradun, 248002, Uttarakhand, India.
Appl Microbiol Biotechnol. 2023 Oct;107(19):6085-6102. doi: 10.1007/s00253-023-12714-x. Epub 2023 Aug 9.
An unprecedented expansion of antifungal therapy failure incidences in healthcare settings of Candida glabrata is the matter of global concern that needs to be addressed efficiently and effectively. In this pursuit, the present study has investigated the antifungal mechanism of benzylisoquinoline alkaloid berberine using biochemical, metabolic, and gene expression analysis, with the aim to delineate its therapeutic activity against C. glabrata and differentially fluconazole-responsive clinical isolates. Interestingly, the clinical isolates were found to be highly susceptible to berberine. Berberine was found to control the surface properties like hydrophobicity and charge of the cells. The cell membrane composition was altered by berberine, where the ergosterol and fatty acids were affected. The efflux pump activity was inhibited, and osmotic stress was generated in C. glabrata cells upon berberine exposure. The berberine has also generated oxidative stress and activated antioxidant system in C. glabrata cells. Furthermore, these observations were supported by the transcriptional expression study of C. glabrata cell genes (CDR1, RLM1, SLT2, SUR4, KRE1) and metabolomics analysis. Based on fold change analysis, the study identified 20 differential metabolites upon berberine treatment, which belong to central carbon, amino acids, and nucleotide pathways. The checkerboard analysis revealed the potentiation of some classically used antifungal drugs by berberine, thus suggesting it as a combinatorial nutraceutical adjuvant for the eradication of fungal infections. KEY POINTS: • Berberine exhibited better potency against azole-resistant clinical isolates • Berberine modulated metabolites of different pathways • Berberine generated oxidative stress and blocked efflux pump activity.
棘白菌素类治疗失败在全球范围内引起了广泛关注，尤其是在假丝酵母菌属中。本研究采用生化、代谢和基因表达分析方法，研究了苯并异喹啉生物碱小檗碱的抗真菌机制，旨在探讨其对假丝酵母菌属和不同氟康唑反应性临床分离株的治疗活性。有趣的是，研究发现临床分离株对小檗碱高度敏感。小檗碱能够控制细胞的表面特性，如疏水性和电荷。细胞膜的组成也因小檗碱而改变，其中麦角固醇和脂肪酸受到影响。小檗碱抑制了细胞的外排泵活性，导致细胞内产生渗透胁迫。小檗碱还能在假丝酵母菌属细胞中产生氧化应激，并激活抗氧化系统。此外，这些观察结果得到了假丝酵母菌属细胞基因（CDR1、RLM1、SLT2、SUR4、KRE1）转录表达研究和代谢组学分析的支持。基于倍数变化分析，研究发现小檗碱处理后有 20 种差异代谢物，这些代谢物属于中心碳、氨基酸和核苷酸途径。棋盘分析显示小檗碱增强了一些经典的抗真菌药物的作用，这表明小檗碱作为一种组合营养辅助剂，可以用于根除真菌感染。关键点：• 小檗碱对唑类耐药的临床分离株表现出更好的活性；• 小檗碱调节了不同途径的代谢物；• 小檗碱产生氧化应激并阻断外排泵活性。
