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400038 重庆,陆军军医大学(第三军医大学)药学与检验医学系微生物与生化药学教研室暨国家免疫生物制品工程技术研究中心
[孙红武, E-mail: sunhongwu2001@163.comSUN Hongwu, E-mail: sunhongwu2001@163.com
[曾浩, E-mail: 023-68771645, zeng1109@163.com ZENG Hao, E-mail: zeng1109@163.com
收稿:2024-05-10,
修回:2024-08-01,
纸质出版:2024-09-30
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赵曼, 吴梓靖, 孙存, 等. 天然植物薰衣草精油对多重耐药鲍曼不动杆菌的抗菌作用及机制研究[J]. 陆军军医大学学报, 2024,46(18):2046-2056.
Man ZHAO, Zijing WU, Cun SUN, et al. Anti-bacterial effect and its mechanism of lavender essential oil against multi-drug resistant
赵曼, 吴梓靖, 孙存, 等. 天然植物薰衣草精油对多重耐药鲍曼不动杆菌的抗菌作用及机制研究[J]. 陆军军医大学学报, 2024,46(18):2046-2056. DOI: 10.16016/j.2097-0927.202405038.
Man ZHAO, Zijing WU, Cun SUN, et al. Anti-bacterial effect and its mechanism of lavender essential oil against multi-drug resistant
目的
2
探讨天然植物薰衣草精油对多重耐药鲍曼不动杆菌的抗菌作用及初步机制。
方法
2
用微量稀释法测定薰衣草精油对多重耐药鲍曼不动杆菌的最低抑菌浓度和最低杀菌浓度;用杀菌动力学研究进一步确定其起效和维持时间;用小鼠感染模型评价薰衣草精油对创口愈合的促进作用;用结晶紫染色定量测定法对其生物膜的抑制和清除作用进行研究;用激光共聚焦显微镜观察生物膜中细菌存活情况;用核酸蛋白测定仪器检测3 mg/mL、6 mg/mL薰衣草精油干预后的细菌DNA核酸和蛋白质泄漏量;用钾离子试剂盒测定钾离子泄漏量,并采用蛋白组学技术结合生物信息学研究方法,初步探究薰衣草精油抗多重耐药鲍曼不动杆菌的作用机制。
结果
2
薰衣草精油的最低抑菌浓度和最低杀菌浓度均为6 mg/mL,在120 min时,该浓度可杀死几乎所有多重耐药鲍曼不动杆菌,且呈现明显的剂量依赖效应和时间依赖效应。整体动物模型评价表明,3 mg/mL和6 mg/mL薰衣草精油均能促进创口愈合,疗效明显。进一步研究发现,3 mg/mL薰衣草精油具有一定的多重耐药鲍曼不动杆菌生物膜抑制作用,且能显著增加DNA核酸和蛋白质的泄漏量,并能促进钾离子外流。同时,6 mg/mL薰衣草精油展现了对多重耐药鲍曼不动杆菌生物膜的清除作用。蛋白组学研究提示,薰衣草精油的抗菌作用可能与影响多重耐药鲍曼不动杆菌的氧化还原酶活性和细胞代谢过程,干扰细胞壁/膜/包膜等结构的生物合成有关。
结论
2
薰衣草精油在3 mg/mL时即可发挥对多重耐药鲍曼不动杆菌的抗菌作用,其作用机制可能与破坏细菌生物膜和干扰细菌代谢有关。
Objective
2
To investigate the antibacterial effect and its preliminary mechanism of lavender essential oil on multi-drug resistant
Acinetobacter baumannii
.
Methods
2
Micro-dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of lavender essential oil against multi-drug resistant
Acinetobacter baumannii
and bactericidal kinetic study was employed to determine the onset and maintenance time of lavender essential oil. Meanwhile
the promoting and therapeutic effects of lavender essential oil on wound healing were observed in a mouse model of infection. Subsequently
crystal violet staining was used to determine the inhibition and clearance of multi-drug resistant
Acinetobacter baumannii
biofilm by lavender essential oil
and laser confocal microscopy was utilized to observe the survival of bacteria in biofilms. NanoDrop instrument was utilized to quantify the leakage of bacterial DNA nucleic acid and protein after intervention with 3 and 6 mg/mL lavender essential oil
and the leakage of bacterial potassium ion was measured by potassium ion test kit. Proteo
mics technology combined with bio-informatics were applied to explore the action mechanism of lavender essential oil against multi-drug resistant
Acinetobacter baumannii
.
Results
2
The MIC and MBC of lavender essential oil were both 6 mg/mL
which could kill almost all multi-drug resistant
Acinetobacter baumannii
at the time point of 120 min
and showed an obvious dose- and time-dependent manner. The overall animal model evaluation showed that both 3 and 6 mg/mL lavender essential oil could promote wound healing
and the curative effect was obvious. Further studies confirmed that 3 mg/mL lavender essential oil had a certain biofilm inhibitory effect on multi-drug resistant
Acinetobacter baumannii
and 6 mg/mL also had a certain biofilm clearance effect under the same conditions. Meanwhile
when incubated at 37 ℃ for 1 h
the dose of 3 mg/mL could increase the leakage of DNA nucleic acid and protein
and significantly promote the efflux of potassium ions. Proteomic analysis suggested that the antibacterial effect of lavender essential oil may be related to affecting the oxidorereductase activity and cell metabolic process of multi-drug resistant
Acinetobacter baumannii
and interfering with the biosynthesis of cell wall/membrane/envelope and other structures.
Conclusion
2
Lavender essential oil at 3 mg/mL can play an antibacterial effect against multi-drug resistant
Acinetobacter baumannii
and its mechanism may be related to the destruction of bacterial biofilm and interference with bacterial metabolism.
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