芦荟大黄素、姜黄素对IncX4型耐药质粒pSD11接合转移的影响  

Effect of aloe-emodin and curcumin on conjugation transfer of IncX4 type resistance plasmid pSD11

作　　者：沈雪 甘德浩 陈璐 郑婷婷 范克伟 邓辉 SHEN Xue;GAN Dehao;CHEN Lu;ZHENG Tingting;FAN Kewei;DENG Hui(College of Animal Sciences,Fujian Agriculture and Forestry University/University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province,Fuzhou,Fujian 350002,China;Fujian Provincial Key Laboratory for Prevention and Control of Animal Infectious Diseases and Biotechnology,Longyan,Fujian 364012,China)

机构地区：[1]福建农林大学动物科学学院/中西兽医结合与动物保健福建省高等学校重点实验室,福建福州350002 [2]福建省家畜传染病防治与生物技术重点实验室,福建龙岩364012

出　　处：《福建农林大学学报(自然科学版)》2025年第3期335-343,共9页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省家畜传染病防治与生物技术重点实验室开放基金课题(2021ZDKF02)。

摘　　要：【目的】研究中药单体芦荟大黄素、姜黄素对IncX4型耐药质粒pSD11接合转移的影响及其相关机制,为了解中药单体影响质粒接合转移效率,降低细菌耐药性提供依据。【方法】通过膜接合转移和平板计数法测定在亚抑菌浓度芦荟大黄素、姜黄素作用下质粒pSD11接合转移的效率。采用实时荧光定量PCR(qRT-PCR)检测接合转移相关基因表达量的变化,并测定中药单体作用下耐药株生物膜的形成能力,分析两种中药单体影响质粒pSD11接合转移的相关机制。【结果】在亚抑菌浓度下,芦荟大黄素、姜黄素能抑制质粒pSD11的接合转移。在抑制接合转移有效浓度下,芦荟大黄素、姜黄素均能上调质粒Ⅳ型分泌系统关键基因的表达量,芦荟大黄素可抑制染色体上接合转移相关基因h-ns、cpxR、pilT的表达量,而不同浓度姜黄素对h-ns、cpxR、pilT的表达量存在多重效应。芦荟大黄素能抑制耐药菌MG1655/pSD11生物膜的形成,而姜黄素对生物膜形成能力的影响总体呈高浓度抑制、低浓度促进的特点。【结论】芦荟大黄素、姜黄素并不是通过抑制Ⅳ型分泌系统关键基因的表达来抑制质粒pSD11的接合转移。芦荟大黄素抑制质粒接合转移的机制包括下调pilT的表达量以及抑制耐药菌生物膜的形成,而不同浓度姜黄素通过影响cpxR、pilT的表达以及生物膜形成抑制质粒接合转移。【Objective】The effects and mechanism of aloe-emodin and curcumin on conjugation transfer of IncX4 type resistance plasmid pSD11 were studied to provide reference for the application of traditional Chinese medicine monomers on inhibiting plasmid conjugation transfer and thus lowering bacterial drug resistance.【Method】The conjugation transfer efficiency of plasmid pSD11 was determined by membrane conjugation transfer and plate counting under aloe-emodin and curcumin at subinhibitory concentration.And their inhibitory mechanism on plasmid conjugation transfer were studied by analyzing the expression of conjugation transfer-related genes through quantitative real-time PCR(qRT-PCR)and biofilm formation ability of resistant strain.【Result】Aloe-emodin and curcumin at the subinhibitory concentration could retard the conjugation transfer of plasmid pSD11.At the effective concentration of inhibiting conjugation transfer,aloe-emodin and curcumin can both up-regulate the expression of key genes in the plasmid typeⅣsecretion system.Aloe-emodin can inhibit the expression of chromosomal conjugation transfer-related gene h-ns,cpxR and pilT,while curcumin exerted multiple effects on their expressions at different concentrations.Furthermore,aloe-emodin can inhibit the biofilm formation of drug-resistance bacteria MG1655/pSD11,while curcumin suppressed biofilm formation at high concentration and facilitated the process at low concentration.【Conclusion】The inhibition of aloe-emodin and curcumin on the conjugation transfer of plasmid pSD11 was not the consequence of regulated expression of key genes involved in typeⅣsecretion system.In general,the inhibitory mechanism of aloe-emodin includes the down-regulation of pilT gene and biofilm inhibition of drug-resistant bacteria,while curcumin at different concentrations acted through multiple effects on cpxR and pilT expressions and biofilm formation.

关 键 词：中药单体 IncX4型质粒 接合转移 细菌生物膜 细菌耐药性 

分 类 号：S853.74[农业科学—临床兽医学]