不同浓度白藜芦醇对高黏液型肺炎克雷伯菌的作用及机制研究
Effect and mechanism of different concentrationsof resveratrol on hypermucinous Klebsiella pneumoniae
俞凤 1徐仙赟 1杨小军 1肖亮 1金桂林 1袁方 1李康 1杨军平1
作者信息
- 1. 江西中医药大学附属医院,江西 南昌 330006
- 折叠
摘要
目的 探讨白藜芦醇(resveratrol)对高黏液型肺炎克雷伯菌(hypermucinous Klebsiella pneumoniae,HMKP)体外生长及生物膜形成的作用及其机制,为开发新型抗菌药物提供初步的理论依据.方法 收集2023年3月—10月江西中医药大学附属医院临床非重复分离HMKP菌株,VITEK-2compact全自动微生物鉴定仪鉴定菌种及对常用抗菌药物的敏感性,采用拉丝试验检测菌株黏液特性,肉汤微量稀释法检测菌株对白藜芦醇最低抑菌浓度(minimal inhibitory concentration,MIC);通过细菌生长曲线法观察不同浓度白藜芦醇对HMKP生长的影响;使用结晶紫染色法测定不同浓度白藜芦醇对HMKP生物膜的影响;应用实时荧光定量(quantitative reverse transcription polymerase chain reaction,RT-qPCR)检测白藜芦醇对HMKP毒力形成相关基因(aerobactin、rmpA和mrkD)的表达量;用GraphPad Prism 9.0绘图并进行数据分析.结果 共收集122株肺炎克雷伯菌,拉丝实验阳性株(HMKP)29株,其中26株对头孢类及其酶抑制剂、喹诺酮类和氨基糖苷类等常用抗菌药物全部敏感,3株为高粘液型耐碳青霉烯类肺炎克雷伯菌(carbapenem-resistant hy-permucinous Klebsiella pneumoniae,HM-CRKP);白藜芦醇对HMKP的MIC均＞512 μg/mL;不同浓度 256 μg/mL,128 μg/mL的白藜芦醇均能明显降低HMKP的生长能力及生物膜形成能力(P＜0.05),且可以降低菌株毒力相关基因rmpA和mrkD的表达量.结论 白藜芦醇能抑制HMKP生长及生物膜形成,其机制可能与通过降低菌株毒力相关基因rmpA和mrkD的表达量有关.
Abstract
Objective To investigate the effects and mechanisms of resveratrol on in vitro growth and biofilm formation of hypermucinous Klebsiella pneumoniae(HMKP),so as to provide a preliminary theoretical basis for the development of new antimicrobial drugs.Methods Clinical isolates of Klebsiella pneumoniae,non-repetitive,collected from March to October 2023,were identified for species and sensitivity to common antimicrobials using the VITEK-2 compact automatic microbial identification system.The mucous characteristics of the strains were assessed via a string test,and the broth microdilution method determined the minimum inhibitory concentration(MIC)of resveratrol.The effects of different concentrations of resveratrol on the growth of hypermucinous Klebsiella pneumoniae were observed by the bacterial growth curve method,and the effects of different concentrations of resveratrol on HMKP biofilm were determined by the crystal violet staining method.Real-time quantitative reverse transcription PCR(RT-qPCR)was used to detect the expression levels of virulence-related genes(aerobactin,rmpA,and mrkD)in HMKP.Data plotting and analysis were performed using GraphPad Prism 9.0.Results A total of 122 strains of Klebsiella pneumoniae were collected,with 29 strains testing positive in the string test(HMKP).Among these,26 strains were allergic to commonly antimicrobial drugs,such as cephalosporins and their enzyme inhibitors,quinolones,and aminoglycosides,while 3 strains were identified as carbapenem-resistant hypermucinous Klebsiella pneumoniae(HM-CRKP).The minimum inhibitory concentration(MIC)of resveratrol for all HMKP strains exceeded 512 μg/mL.Resveratrol concentrations of 256 μg/mL and 128 μg/mL significantly reduced the growth capacity and biofilm formation capacity of HMKP(P＜0.05)and decreased the expression levels of virulence-related genes rmpA and mrkD.Conclusions Resveratrol can inhibit the growth and biofilm formation of HMKP.The mechanism may be related to the downregulation of virulence-related genes expression,specifically rmpAandmrkD.
关键词
白藜芦醇/高黏液型肺炎克雷伯菌/生物膜/毒力基因Key words
Resveratrol/hypermucinous Klebsiella pneumoniae/biofilms/virulence genes引用本文复制引用
基金项目
江西省中医药局科技项目(2019A076)
江西省卫生健康委科技项目(202210796)
出版年
2024
NETL
NSTL
万方数据