肉桂醛与麝香草酚对金黄色葡萄球菌的协同抑菌活性及作用机制

doi:10.16431/j.cnki.1671-7236.2024.12.039

摘要/Abstract

摘要： 【目的】探究肉桂醛与麝香草酚联合作用对金黄色葡萄球菌的抑菌活性及作用机制，开发新型植物精油抗菌剂，为预防和治疗奶牛乳腺炎临床药物的应用提供理论依据。【方法】本研究采用微量棋盘稀释法测定肉桂醛与麝香草酚的联合抑菌指数；利用时间-杀菌曲线进一步验证协同杀菌能力，通过观察金黄色葡萄球菌膜完整性、膜通透性、细胞膜电位及测定生物膜形成量、碱性磷酸酶(AKP)活性探究肉桂醛和麝香草酚对细菌壁膜结构的影响；同时研究肉桂醛和麝香草酚对菌体活性氧(ROS)生成的影响。【结果】联合药敏试验结果显示，肉桂醛与麝香草酚联用时表现出良好的协同抑菌活性，联合抑菌指数在0.1875～0.375之间。体外时间-杀菌曲线结果进一步表明，联合用药组可明显降低细菌数量。肉桂醛和麝香草酚协同作用可降低细胞膜完整性，显著增加细胞膜通透性(P＜0.05)，细胞膜电位去极化，显著降低生物膜形成量(P＜0.05)，显著增加金黄色葡萄球菌AKP外渗量(P＜0.05)，显著提高ROS水平(P＜0.05)，进而导致细胞损伤和死亡。【结论】肉桂醛与麝香草酚协同作用通过诱导细胞膜损伤，导致细胞内容物泄漏及破坏细胞内稳态发挥抗菌活性。本研究可为开发治疗奶牛乳腺炎的天然植物精油抗菌剂肉桂醛和麝香草酚的临床应用提供参考。

关键词: 肉桂醛; 麝香草酚; 金黄色葡萄球菌; 协同抑菌活性

Abstract: 【Objective】 This study was aimed to explore the antibacterial activity and mechanism of the combined action of cinnamaldehyde (CA) and thymol (Thy) on Staphylococcus aureus (S.aureus),develop a novel plant-derived essential oil antibacterial agent,establish a theoretical foundation for the clinical application in preventing and treating mastitis in dairy cows.【Method】 In this study,the fractional inhibitory concentration index (FICI) of CA and Thy was determined using the micro-checkerboard dilution method.The time-sterilization curve was further used to verify the synergistic bactericidal ability.The effects of CA and Thy on the bacterial cell membrane structure were studied by observing the membrane integrity,membrane permeability,cell membrane potential,biofilm formation,and alkaline phosphatase (AKP) activity of S.aureus.The effect of CA and Thy on reactive oxygen species (ROS) production was also examined.【Result】 CA and Thy demonstrated significant synergistic antibacterial activity,with a FICI between 0.1875 and 0.375. The results of the time-killing test confirmed the synergistic effect of the CA and Thy combination on S.aureus.The cell membrane integrity significantly decreased,the cell membrane permeability significantly increased (P＜0.05),the cell membrane potential depolarized,and the biofilm formation significantly decreased under the synergistic effect of CA and Thy (P＜0.05).The synergistic effect of CA and Thy significantly increased the AKP extravasation of S.aureus (P＜0.05) and significantly increased ROS level (P＜0.05),leading to cell damage and death.【Conclusion】 In summary,the synergistic combination of CA and Thy exerted antibacterial activity by inducing cell membrane damage,leading to leakage of cell contents and disruption of intracellular homeostasis.This study provided a reference for the clinical application of the natural plant essential oil antibacterial agents CA and Thy in the treatment of mastitis in dairy cows.

Key words: cinnamaldehyde; thymol; Staphylococcus aureus; synergistic antibacterial activity

中图分类号:

S859.7

陈晓慧, 王桂琴, 王建崇, 何小强, 陈昊蓉. 肉桂醛与麝香草酚对金黄色葡萄球菌的协同抑菌活性及作用机制[J]. 中国畜牧兽医, 2024, 51(12): 5528-5538.

CHEN Xiaohui, WANG Guiqin, WANG Jianchong, HE Xiaoqiang, CHEN Haorong. Synergistic Antibacterial Activity and Mechanism of Cinnamaldehyde and Thymol Against Staphylococcus aureus[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5528-5538.

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