异丙氧苯胍联合黏菌素对肺炎克雷伯菌体外协同抗菌作用

doi:10.11843/j.issn.0366-6964.2024.12.041

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (12): 5792-5801.doi: 10.11843/j.issn.0366-6964.2024.12.041

异丙氧苯胍联合黏菌素对肺炎克雷伯菌体外协同抗菌作用

吴素娟¹^,²^,³(), 林昌成¹^,²^,³, 万鹏¹^,²^,³, 李杰¹^,²^,³, 陆毅兴¹^,²^,³, 胡健欣¹^,²^,³, 彭险峰⁴, 曾振灵¹^,²^,³^,*()

1. 华南农业大学兽医学院, 广州 510642
2. 广东省兽药研制与安全评价重点实验室, 广州 510642
3. 国家兽医微生物耐药性风险评估实验室, 广州 510642
4. 广东英赛特生物技术有限公司, 广州 510000

收稿日期:2024-01-26 出版日期:2024-12-23 发布日期:2024-12-27
通讯作者: 曾振灵 E-mail:2411924405@qq.com;zlzeng@scau.edu.cn
作者简介:吴素娟(1999-), 女, 福建宁德人, 硕士, 主要从事兽医药理学与毒理学研究, E-mail: 2411924405@qq.com
基金资助:
国家自然科学基金项目(32273057)

Synergistic Antimicrobial Effect of Isopropoxy Benzene Guanidine Combined with Colistin on Klebsiella pneumoniae in vitro

WU Sujuan¹^,²^,³(), LIN Changcheng¹^,²^,³, WAN Peng¹^,²^,³, LI Jie¹^,²^,³, LU Yixing¹^,²^,³, HU Jianxin¹^,²^,³, PENG Xianfeng⁴, ZENG Zhenling¹^,²^,³^,*()

1. College of Veterinary Medicine, South China Agriculture University, Guangzhou 510642, China
2. Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety, Guangzhou 510642, China
3. National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, Guangzhou 510642, China
4. Guangzhou Insighter Biotechnology Co. Ltd, Guangzhou 510000, China

Received:2024-01-26 Online:2024-12-23 Published:2024-12-27
Contact: ZENG Zhenling E-mail:2411924405@qq.com;zlzeng@scau.edu.cn

摘要/Abstract

摘要：

旨在研究异丙氧苯胍与黏菌素的协同抗菌能力及初步联合作用机制，评估异丙氧苯胍是否能作为一种有效的黏菌素增效剂，增强黏菌素抗菌效果，以期为临床上多重耐药肺炎克雷伯菌感染提供有效治疗的理论依据。本研究通过异丙氧苯胍与黏菌素对肺炎克雷伯菌的最低抑菌浓度、棋盘试验、时间杀菌曲线、耐药性诱导和荧光染料试验对协同抗菌及初步联合机制进行研究。结果显示：异丙氧苯胍单用对肺炎克雷伯菌没有抗菌活性，但与黏菌素联用可对肺炎克雷伯菌产生协同作用，与阿莫西林联用时具有相加作用。时间杀菌曲线表明，两药联合使用时，异丙氧苯胍可增强肺炎克雷伯菌对黏菌素的敏感性，细菌数量与黏菌素单药作用相比下降＞2 lg(CFU·mL^-1)，具有协同作用。在长期诱导耐药性试验中，较黏菌素单药组、联合用药组中黏菌素更不易诱导产生耐药性。协同作用机制试验结果表明，黏菌素与异丙氧苯胍联合使用时，可以破坏细胞质膜通透性，影响细胞膜电势，消散质子动力势能，使细胞膜难以发挥正常功能。本研究发现异丙氧苯胍与低浓度黏菌素联用对肺炎克雷伯菌具有良好协同抗菌活性，增强黏菌素的抗菌作用。综上表明，异丙氧苯胍可以恢复多重耐药肺炎克雷伯菌对黏菌素的敏感性，可作为治疗肺炎克雷伯菌感染的潜在治疗方案。

关键词: 肺炎克雷伯菌, mcr, 异丙氧苯胍, 黏菌素, 协同抗菌活性

Abstract:

This work aimed to study the antibacterial synergistic ability and the preliminary combined mechanism of isopropoxy benzene guanidine (IBG) combined with colistin, and to evaluate whether IBG can act as an effective colistin synergist and enhance the anti-bacterial effect of colistin. The findings of this study will provide a theoretical basis for the effective treatment of multidrug resistant bacterial infections in the clinic. This study investigated the preliminary combined mechanism and synergistic effect of IBG with colistin against Klebsiella pneumoniae using the minimal inhibitory concentration, checkerboard test, time killing curves, resistance induction and fluorescent dye test. IBG has no antimicrobial activity against K. pneumoniae alone, but has a synergistic effect against K. pneumoniae in combination with colistin and an additive effect in combination with amoxicillin. Time killing curves indicate that when the two drugs are used in combination, IBG enhances the susceptibility of K. pneumoniae to colistin, with a synergistic decrease in bacterial counts of >2 lg(CFU·mL^-1) compared to the effect of colistin alone. In the long-term induced resistance test, resistance was less likely to be induced by colistin in the combination group than in the colistin monotherapy group. The results of the synergistic mechanism test showed that colistin and IBG, when used in combination, can destroy the permeability of the cytoplasmic membrane, affect the potential of the cell membrane, and dissipate the proton kinetic potential energy, so that it is difficult for cell membranes to perform their normal functions. In this study, we found that the combination of IBG and low concentration of colistin, has good synergistic antibacterial activity against K. pneumoniae and enhances the antibacterial effect of colistin. It suggests that IBG can restore the susceptibility of multidrug resistant K. pneumoniae to colistin, and can be used as a potential therapeutic option for the treatment of K. pneumoniae infections.

Key words: Klebsiella pneumoniae, mcr, isopropoxy benzene guanidine, colistin, synergistic antimicrobial activity

中图分类号:

S859.796

吴素娟, 林昌成, 万鹏, 李杰, 陆毅兴, 胡健欣, 彭险峰, 曾振灵. 异丙氧苯胍联合黏菌素对肺炎克雷伯菌体外协同抗菌作用[J]. 畜牧兽医学报, 2024, 55(12): 5792-5801.

WU Sujuan, LIN Changcheng, WAN Peng, LI Jie, LU Yixing, HU Jianxin, PENG Xianfeng, ZENG Zhenling. Synergistic Antimicrobial Effect of Isopropoxy Benzene Guanidine Combined with Colistin on Klebsiella pneumoniae in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5792-5801.

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菌株 Strain	MIC/(μg·mL^-1)
菌株 Strain	IBG	CL
ATCC 700603	>1 280	1
P135	>1 280	4
C14	>1 280	8
C20	>1 280	16
124C	>1 280	16

联用组合 Combination	MIC/(μg·mL^-1)		部分抑菌浓度指数 FICI	作用关系 Functional relationship
联用组合 Combination	异丙氧苯胍联合/单药 IBG(in combination)/IBG	B药联合/单药 Drug B(in combination)/Drug B	部分抑菌浓度指数 FICI	作用关系 Functional relationship
IBG+CL	64/1 000	0.25/1.00	0.31	协同
IBG+AMO	8/1 000	32/64	0.50	相加
IBG+TMP	32/1 000	32/32	1.03	无关
IBG+GEN	32/1 000	0.5/8.0	0.10	无关
IBG+AMP	4/1 000	8/4	2.00	无关
IBG+CAZ	256/1 000	16/16	1.25	无关
IBG+MEM	4/1 000	0.5/0.5	1.00	无关
IBG+FOX	256/1 000	32/32	1.25	无关
IBG+TIG	4/1 000	2/1	2.00	无关
IBG+CIP	256/1 000	64/64	1.25	无关
IBG+FFC	8/1 000	32/16	2.00	无关
IBG+NEO	8/1 000	4/2	2.00	无关

菌株 Strains	耐药基因 Resistance genes	MIC/(μg·mL^-1)		部分抑菌浓度指数 FICI	作用关系 Functional relationship
菌株 Strains	耐药基因 Resistance genes	异丙氧苯胍联合/单药 IBG(in combination)/IBG	黏菌素联合/单药 CL(in combination)/CL	部分抑菌浓度指数 FICI	作用关系 Functional relationship
ATCC 700603	-	64/1 000	0.25/1.00	0.31	协同
P135	bla_SHV、mcr-8	32/1 000	0.25/4.00	0.10	协同
124C	bla_CTX-M、bla_SHV	32/1 000	0.125/16.000	0.04	协同
C14	bla_SHV	32/1 000	0.5/8.0	0.10	协同
C20	bla_SHV	32/1 000	0.125/16.000	0.04	协同

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异丙氧苯胍联合黏菌素对肺炎克雷伯菌体外协同抗菌作用

Synergistic Antimicrobial Effect of Isopropoxy Benzene Guanidine Combined with Colistin on Klebsiella pneumoniae in vitro

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