The emergence of resistant Candida albicans has made clinical fluconazole (FLC) treatment difficult. Improving sensitivity to FLC is an effective way to treat resistant isolates. Berberine hydrochloride (BBH) is a commonly used traditional Chinese medicine with antimicrobial effects, especially in resistant isolates. We investigated the molecular mechanisms underlying BBH and FLC synergism on biofilm-positive FLC-resistant C. albicans inhibition. Checkerboard microdilution assays and time-kill assays showed a strong synergistic effect between BBH and FLC in resistant C. albicans isolates, causing a significant 32–512-fold reduction in minimum inhibitory concentrations. BBH combined with FLC inhibited intracellular FLC efflux due to key efflux pump gene CDR1 downregulation, whereas FLC alone induced high CDR1 transcription in resistant strains. Further, BBH + FLC inhibited yeast adhesion, morphological hyphae transformation, and biofilm formation by downregulating the hyphal-specific genes ALS3, HWP1, and ECE1. BBH caused cytoplasmic Ca²⁺ influx, while FLC alone did not induce high intracellular Ca²⁺ levels. The vacuolar calcium channel gene YVC1 was upregulated, while the vacuolar calcium pump gene PMC1 was downregulated in the BBH + FLC and BBH alone groups. However, vacuolar calcium gene expression after FLC treatment was opposite in biofilm-positive FLC-resistant C. albicans, which might explain why BBH induces Ca²⁺ influx. These results demonstrate that BBH + FLC exerts synergistic effects to increase FLC sensitivity by regulating multiple targets in FLC-resistant C. albicans. These findings further show that traditional Chinese medicines have multi-target antimicrobial effects that may inhibit drug-resistant strains. This study also found that the vacuolar calcium regulation genes YVC1 and PMC1 are key BBH + FLC targets which increase cytoplasmic Ca²⁺ in resistant isolates, which might be critical for reversing biofilm-positive FLC-resistant C. albicans.

抗药性的出现 白色念珠菌已经使氟康唑（FLC）的临床治疗变得困难。提高对FLC的敏感性是治疗耐药菌株的有效方法。盐酸小ber碱（BBH）是一种具有抗菌作用的常用中药，尤其是在耐药菌中。我们调查了BBH和FLC协同作用对生物膜阳性FLC耐药的潜在分子机制白色念珠菌抑制。棋盘微稀释测定法和时间杀灭测定法显示BBH和FLC在抗药性方面具有很强的协同作用白色念珠菌分离物，导致最低抑菌浓度显着降低32–512倍。BBH结合FLC抑制了细胞内FLC流出，这是由于关键的流出泵基因CDR1 下调，而单独的FLC诱导高 CDR1抗性菌株中的转录。此外，BBH + FLC通过下调菌丝特异性基因来抑制酵母菌粘附，形态菌丝转化和生物膜形成ALS3， HWP1和 ECE1。BBH引起细胞质Ca ²⁺大量涌入，而单独的FLC不能诱导高的细胞内Ca ²⁺水平。液泡钙通道基因YVC1 被上调，而液泡钙泵基因 PMC1在BBH + FLC和BBH单独组中被下调。然而，FLC处理后液泡钙基因表达在生物膜阳性FLC耐药中相反白色念珠菌，这可能解释了BBH为何诱导Ca ^2+内流。这些结果表明，BBH + FLC通过调节抗FLC的多个靶点发挥协同作用，从而提高FLC敏感性。白色念珠菌。这些发现进一步表明，中药具有多靶点抗菌作用，可能会抑制耐药菌株。这项研究还发现液泡钙调节基因YVC1 和 PMC1是重要的BBH + FLC靶标，可增加抗性分离物中的细胞质Ca ²⁺，这对于逆转生物膜阳性的FLC耐药性可能至关重要白色念珠菌。