聚苯乙烯微塑料对肺炎克雷伯菌生长、生物膜形成和毒力的影响  

Effects of polystyrene microplastics on the growth,biofilm formation and virulence of Klebsiella pneumoniae

作　　者：程振富 白淼 白宇超 赵乾秀 王正豪 张灿 张传福 Cheng Zhenfu;Bai Miao;Bai Yuchao;Zhao Qianxiu;Wang Zhenghao;Zhang Can;Zhang Chuanfu(Dept of Health Inspection and Quarantine,School of Public Health,Anhui Medical University,Hefei 230032;Chinese People's Liberation Army Center for Disease Control and Prevention,Beijing 100071)

机构地区：[1]安徽医科大学公共卫生学院卫生检验与检疫学系,合肥230032 [2]中国人民解放军疾病预防控制中心,北京100071

出　　处：《安徽医科大学学报》2024年第10期1835-1841,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家重点研发计划项目(编号:2023YFF0614204);国家自然科学基金项目(编号:52070193)。

摘　　要：目的探究聚苯乙烯微塑料(PS-MPs)对肺炎克雷伯菌生长、活性、氧化应激水平、生物膜形成和毒力的影响。方法肺炎克雷伯菌暴露在不同浓度(10、50和100μg/ml)和粒径(0.1、1.0和5.0μm)的PS-MPs环境下,测定生长曲线;通过CCK-8测定细菌活性;荧光探针法测定胞内活性氧(ROS)水平;结晶紫染色法测定生物膜形成能力;使用实时荧光定量PCR(qRT-PCR)检测生物膜形成相关基因(luxS、mrkA、wbbM、pgaA、wzm)和毒力基因(ureA、uge、wabG、fimH)的相对表达水平。结果高浓度(100μg/ml)的0.1μm PS-MPs对肺炎克雷伯菌的生长和活性具有更强的抑制作用,胞内ROS水平显著提升,表明粒径越小、浓度越高的PS-MPs对细菌的毒性越强。100μg/ml各粒径组(0.1、1.0和5.0μm)的PS-MPs显著促进肺炎克雷伯菌的生物膜形成,提高生物膜形成相关基因(luxS、mrkA、wbbM、pgaA、wzm)和毒力基因(ureA、uge、wabG、fimH)的相对表达水平。结论PS-MPs通过诱导肺炎克雷伯菌产生较高水平ROS,引起氧化应激,导致细菌的生长和活性受到抑制,并且增强生物膜形成能力和毒力,从而影响肺炎克雷伯菌的生物学特性。Objective To explore the effects of polystyrene microplastics(PS-MPs)on the growth,activity,oxidative stress levels,biofilm formation and virulence of Klebsiella pneumoniae.Methods Klebsiella pneumoniae was exposed to PS-MPs at different concentrations(10,50 and 100μg/ml)and particle sizes(0.1,1.0 and 5.0μm),and the growth curves were measured.The bacterial activity was determined by CCK-8(cell counting kit-8).The level of intracellular reactive oxygen species(ROS)was determined by fluorescence probe.The biofilm forming ability was determined by crystal violet staining.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the relative expression levels of biofilm-forming genes(luxS,mrkA,wbbM,pgaA,wzm)and virulence genes(ureA,uge,wabG,fimH).Results A high concentration(100μg/ml)of 0.1μm PS-MPs had a stronger inhibitory effect on the growth and activity of Klebsiella pneumoniae,and the intracellular ROS level significantly increased,indicating that smaller particle size and higher concentration of PS-MPs were more toxic to bacteria.PS-MPs of 100μg/ml particle size groups(0.1,1.0 and 5.0μm)significantly promoted the biofilm formation of Klebsiella pneumoniae.The relative expression levels of biofilm formation related genes(luxS,mrkA,wbbM,pgaA,wzm)and virulence genes(ureA,uge,wabG,fimH)increased.Conclusion By inducing Klebsiella pneumoniae to produce a high level of ROS,PS-MPs can cause oxidative stress,inhibit the growth and activity of bacteria,and enhance the biofilm formation ability and virulence,thus affecting the biological characteristics of Klebsiae pneumoniae.

关 键 词：微塑料 肺炎克雷伯菌 氧化应激 生物膜形成 毒力 

分 类 号：R114[医药卫生—卫生毒理学]