大蒜E素对副溶血弧菌的抑菌活性及机制

ANTIBACTERIAL ACTIVITY AND MECHANISM OF ALLICIN E AGAINST VIBRIO PARAHAEMOLYTICUS

  • 摘要: 为探究大蒜E素(ALE)对副溶血弧菌的抑菌活性及作用机制, 通过测定最小抑菌浓度(MIC)、最小杀菌浓度(MBC)和抑菌动态生长曲线, 评价ALE对副溶血弧菌的抑菌作用; 基于电镜观察、β-D-半乳糖苷酶活性及蛋白质泄漏量, 分析其对副溶血弧菌细胞结构的影响; 采用结晶紫染色法检测ALE对副溶血弧菌生物被膜形成的影响; 利用RNA-Seq技术分析ALE对副溶血弧菌转录水平表达的影响。结果显示, ALE对副溶血弧菌的MIC为7.8 μg/mL, MBC为31.2 μg/mL。与对照组相比, MIC浓度ALE组副溶血弧菌胞外β-D-半乳糖苷酶活性和蛋白质含量显著增加(P<0.05), 细胞壁膜消融、内容物流失, 生物被膜形成量显著减少(P<0.05)。转录组分析结果显示, 与对照组相比, 1/4MIC浓度的ALE组副溶血弧菌中显著下调基因145个, 主要富集在MAPK信号转导、鞭毛组装、生物膜形成等通路; ALE组副溶血弧菌中显著上调基因133个, 主要富集在氨基酸代谢、降解等通路; RNA-Seq结果与RT-qPCR验证试验结果一致。研究表明, ALE对副溶血弧菌抑菌作用强, 可能通过破坏细菌菌体结构, 抑制其生物被膜形成, 以及调控鞭毛、生物膜等基因表达, 从而发挥抑菌、减毒作用。研究结果为探明ALE抑制副溶血弧菌的作用及机理奠定良好的基础。

     

    Abstract: To investigate the antibacterial activity of Allicin E (ALE) against Vibrio parahaemolyticus, the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and growth curve were determined. The antibacterial mechanisms were investigated by exploring the influence of ALE on the extracellular β-D-galactosidase activity, protein content, bacterial morphological structure, biofilm formation ability, and transcriptome of V. parahaemolyticus. The results showed that the MIC and MBC of ALE against V. parahaemolyticus were 7.8 and 31.2 μg/mL, respectively. Treatment with ALE resulted in damage to the cell wall and membrane structure of V. parahaemolyticus, leading to the leakage of intracellular substances. Compared with the control group, extracellular β-D-galactosidase activity and protein content in the ALE-treated group were significantly increased (P<0.05), while the amount of bacterial biofilm formation was significantly decreased (P<0.05). The transcriptome analysis results showed that 145 genes were significantly downregulated in the ALE group, mainly enriched in pathways such as MAPK signal transduction, flagella assembly, and biofilm formation in Pseudomonas aeruginosa. Conversely, 133 genes were significantly upregulated, mainly related to amino acid metabolism and degradation pathways. The RNA-seq results were consistent with the RT-qPCR validation tests. Our results demonstrated that ALE showed a strong antimicrobial effect against V. parahaemolyticus by disrupting bacterial structure, inhibiting biofilm formation, and regulating the expression of genes such as flagella and biofilms. This study laid an important foundation for exploring the role and mechanism of ALE against V. parahaemolyticus.

     

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