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<p>Lupinifolin possesses the capability to combat specific pathogenic bacteria. Nonetheless, Gram-negative bacteria exhibit minimal inhibition by lupinifolin. This study evaluated the synergistic antibacterial and antibiofilm activities of combinations of lupinifolin and antibiotics against pathogenic bacteria using a checkerboard assay and time-kill analysis. Crystal violet staining and confocal laser scanning microscopy (CLSM) were utilized to evaluate antibiofilm properties. After treatment, gene expression changes in isolates were evaluated using quantitative real-time PCR. Lupinifolin and streptomycin synergistically inhibited MRSA and VRE with a FIC index of 0.5, whereas lupinifolin and vancomycin similarly inhibited <i>S. aureus</i> ATCC25923. Partially synergistic interaction against <i>E. faecalis</i> ATCC29212 between lupinifolin and tetracycline, vancomycin, and chloramphenicol with an FIC index of 0.625, 0.5625, and 0.625, respectively. Partially synergistic interaction against <i>E. faecium</i> HTY0256 between lupinifolin and tetracycline with an FIC index of 0.625. The interaction between lupinifolin and streptomycin against Gram-negative bacteria, such as <i>E. coli</i> ATCC25922, showed a synergistic effect. The time-kill assay of the combinations showed an inhibitory effect of more than 3 log and 2 log (CFU/ml) at 4&#xa0;h incubation. CLSM revealed that the combinations reduced biofilm formation. The combination of lupinifolin and streptomycin altered biofilm-forming genes in <i>E. faecalis</i> ATCC29212 and VRE isolates; <i>gelE</i> and <i>bepA</i> were down-regulated. In contrast, MRSA isolates had <i>sarA</i> and <i>ebpS</i> up-regulated. In addition, our findings suggested that lupinifolin destroys cell membranes, as demonstrated by the expression of <i>secA</i>. Our investigation enabled the prospective integration of lupinifolin with antibiotics, enhancing their efficacy and application against antimicrobial-resistant pathogens.</p>

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Synergistic antibacterial and antibiofilm properties of lupinifolin in combination with antibiotics against pathogenic bacteria

  • 期刊论文

  • 发表日期：

    2025年11月20日

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  • Sutthisa Khwankaew,
  • Chutimon Promthong,
  • Nantiya Joycharat,
  • Mingkwan Yingkajorn,
  • Wipawadee Sianglum

摘要

Lupinifolin possesses the capability to combat specific pathogenic bacteria. Nonetheless, Gram-negative bacteria exhibit minimal inhibition by lupinifolin. This study evaluated the synergistic antibacterial and antibiofilm activities of combinations of lupinifolin and antibiotics against pathogenic bacteria using a checkerboard assay and time-kill analysis. Crystal violet staining and confocal laser scanning microscopy (CLSM) were utilized to evaluate antibiofilm properties. After treatment, gene expression changes in isolates were evaluated using quantitative real-time PCR. Lupinifolin and streptomycin synergistically inhibited MRSA and VRE with a FIC index of 0.5, whereas lupinifolin and vancomycin similarly inhibited S. aureus ATCC25923. Partially synergistic interaction against E. faecalis ATCC29212 between lupinifolin and tetracycline, vancomycin, and chloramphenicol with an FIC index of 0.625, 0.5625, and 0.625, respectively. Partially synergistic interaction against E. faecium HTY0256 between lupinifolin and tetracycline with an FIC index of 0.625. The interaction between lupinifolin and streptomycin against Gram-negative bacteria, such as E. coli ATCC25922, showed a synergistic effect. The time-kill assay of the combinations showed an inhibitory effect of more than 3 log and 2 log (CFU/ml) at 4 h incubation. CLSM revealed that the combinations reduced biofilm formation. The combination of lupinifolin and streptomycin altered biofilm-forming genes in E. faecalis ATCC29212 and VRE isolates; gelE and bepA were down-regulated. In contrast, MRSA isolates had sarA and ebpS up-regulated. In addition, our findings suggested that lupinifolin destroys cell membranes, as demonstrated by the expression of secA. Our investigation enabled the prospective integration of lupinifolin with antibiotics, enhancing their efficacy and application against antimicrobial-resistant pathogens.