重组人β-防御素3对铜绿假单胞菌生物膜结构影响的初步研究  

Inhibitory effects of recombinant human beta-defensin 3 on biofilm structure of Pseudomonas aeruginosa:a preliminary study in vitro

作　　者：周菁[1] 陈章[1] 田坤[1] 吴奎[1] 肖贞良[1] 

机构地区：[1]成都军区总医院呼吸科,成都610083

出　　处：《西南国防医药》2011年第1期4-7,共4页Medical Journal of National Defending Forces in Southwest China

基　　金：全军"十一五"课题面上项目(06MB191);成都军区"十一五"课题面上项目(MB07015)

摘　　要：目的研究重组人β-防御素3(rhBD-3)对铜绿假单胞菌(PA)形成生物膜的体外抑制作用。方法平板培养法培养PA,得到早期和成熟期生物膜;琼脂糖弥散抗菌法测定最低抑菌浓度(M IC);扫描电镜(SEM)观察载体表面BF形态;连续稀释法行活菌计数。结果 rhBD-3对PA的M IC值是64μg/m l。rhBD-3作用后8、16、24 h,膜片载体表面PA活菌黏附量减少,且随着rhBD-3浓度的增高,活菌群数量进行性减少;SEM观察见rhBD-3组少量散在的游离细菌,有少量小斑片状多糖复合物,而空白对照组可见大量分布均匀的微菌落和游离细菌及多糖复合物交联。结论 rhBD-3可以抑制PA的BF形成,并对早期和成熟期BF具有破坏作用。Objective To investigate the inhibitory effects of recombinant human beta-defensin 3（rhBD-3）on the biofilm formation of Pseudomonas aeruginosa（PA） in vitro.Methods Standard gelose radial diffusion assay was used to measure the minimal inhibitory concentration（MIC） of rhBD-3 on PA.Then the early and mature biofilms of PA inoculated on polycarbonate membrane were obtained using plate culture method.These BFs were randomly divided into four groups：blank control group and three groups treated with different concentration of rhBD-3（1/2MIC,2MIC and 4MIC,respectively）.The appearance of biofilms on vehicle surface was observed under scanning electron microscope（SEM）.Viable count was conducted by serial dilution method.Results The MIC of rhBD-3 on Pseudomonas aeruginosa was 64 μg/ml.The amount of viable bacterial adhered to the vehicle surface decreased 8,16 and 24 h after rhBD-3 treatment.Furthermore,it decreased progressively with the increase of rhBD-3 concentration.Small amounts of sporadic and free bacteria as well as patchy polyose compounds were seen in rhBD-3 groups under SEM.However,large amounts of evenly distributed microcolony and free bacteria as well as the crosslinking of polyose compounds were found in blank control group.Conclusion rhBD-3 can prevent PA from forming BF in vitro.It can also destroy the structure of this biofilm in early or mature stage.

关 键 词：铜绿假单胞菌 生物膜 人β-防御素3 琼脂糖弥散抗菌法 

分 类 号：R378[医药卫生—病原生物学]