不同氧浓度对铜绿假单胞菌生长和毒力的调节作用  

The effect of different oxygen concentrations regulation the growth and virulence of Pseudomonas aeruginosa

作　　者：陈双红[1] 陈锐勇[1] 徐雄利[1] 田力[1] 

机构地区：[1]中国人民解放军海军医学研究所舰艇卫生研究室,高气压与潜水生理学全军重点实验室,上海200433

出　　处：《职业与健康》2017年第22期3057-3060,3065,共5页Occupation and Health

基　　金：国家自然基金面上项目(81271796)

摘　　要：目的研究不同氧浓度对铜绿假单胞菌生长及毒力的调节作用,阐明氦氧饱和高气压潜水富氧环境中铜绿假单胞菌感染高发机制。方法以铜绿假单胞菌PAO1(ATCC15692)为研究对象,主要设置实验对照组(20%O_2)、富氧实验组(40%O_2)、低氧实验组(2%O_2)共三个暴露组。用半固体培养基、丛集运动培养基观察细菌生长及运动能力,绿脓菌素测定培养基结合光密度法测定绿脓菌素含量。用HE染色结合CCK-8方法,观察细菌培养上清对靶细胞A549的损伤效应。结果与对照组相比,富氧环境生长的细菌克隆厚、中心明显隆起,运动生长轨迹不明显;低氧环境生长的细菌克隆中心凹陷、平铺生长,细菌运动生长轨迹显著。相较于实验对照组,低氧实验组绿脓菌素分泌能力降低(0.38±0.08 vs 0.16±0.03,F=10.22,P=0.033),富氧浓度组绿脓菌素分泌能力增加(0.38±0.08 vs 0.66±0.06,F=24.00,P=0.008)。细胞毒性检测显示,与空白对照组(F=93.55,P=0.001)和实验对照组(F=14.69,P=0.019)相比,富氧暴露生长细菌分离上清与细胞共培养8 h后,靶细胞出现明显损伤效应。结论富氧浓度可通过调节铜绿假单胞菌生长、运动及毒力,诱导铜绿假单胞菌致病能力。[Objective]To study the regulation effect of different oxygen concentration on the growth and toxicity of pseudomonas aeruginosa,and illuminate the high epidemic mechanism of pseudomonas aeruginosa infection in helium oxygen saturation high pressure diving environment. [Methods]Pseudomonas aeruginosa（ATCC15692） was as study object,three exposure groups including experimental control group（20% O_2）,oxygen enriched experimental group（40% O_2）and low oxygen experimental group（2% O_2）were set. Semi-solid medium and swarming medium were used to observe the growth and motility of bacteria.Pyocyanin determination medium combined with optical density determination method were used to detect the concentration of pyocyanin. HE staining combined with CCK-8 method was used to observe the injury effect of bacterial supernatant on A549 cell.[Results]Compared with control group,the bacteria grew in oxygen enriched environment had thick clone,obviously ridgy center and non obvious movement growth track,and the bacteria grew in low oxygen environment had sunk clone center,flat growth and obvious movement growth track. Compared with the experimental control group,secretion ability of pseudomonas aeruginosa in low oxygen experimental group decreased（0.38±0.08 vs 0.16±0.03,F=10.22,P=0.033）,and increased in oxygen enriched experimental group（0.38 ±0.08 vs 0.66 ±0.06,F =24.00,P =0.008）. Cytotoxicity test showed that compared with blank control group（F=93.55,P=0.001）and experimental control group（F=14.69,P=0.019）,the target cells were obviously damaged after separated supernatant and cells were cultured in oxygen enriched environment for 8 h. [Conclusion]The concentration of enriched oxygen can induce the pathogenicity of pseudomonas aeruginosa by regulating its growth,movement and toxicity.

关 键 词：铜绿假单胞菌 氧 暴露 绿脓菌素 毒力 

分 类 号：R117[医药卫生—公共卫生与预防医学]