亚胺培南/西司他丁对铜绿假单胞菌生物膜形成和藻酸盐合成基因表达的影响  

Effect of Imipenem/cilastatin on Biofilm Formation and Alginate Biosynthesis Gene Expression of Pseudomonas Aeruginosa

作　　者：倪明[1] 余冰[2] 田德英[1] 宋世会[1] 谢琳卡[1] 陈安群[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院,武汉430030 [2]华中科技大学同济医学院病原生物学系

出　　处：《内科急危重症杂志》2008年第4期190-193,共4页Journal of Critical Care In Internal Medicine

基　　金：国家自然科学基金资助项目(No:30571645);湖北省自然科学基金资助项目(No:2007ABA177)

摘　　要：目的:研究亚胺培南/西司他丁对黏液型铜绿假单胞菌PA17及非黏液型铜绿假单胞菌PAO1生物膜形成及其藻酸盐合成基因algD表达的影响。方法:试管倍比稀释法检测亚胺培南/西司他丁对PA17和PAO1的最低抑菌浓度(MIC);改良平板培养法建立PA17和PAO1的生物膜模型,从微菌落形成阶段开始在生物膜培养基中分别加入1倍和10倍最低抑菌浓度的亚胺培南/西司他丁,扫描电镜观察PA17和PAO1生物膜形成的变化,半定量RT-PCR检测加入亚胺培南/西司他丁对生物膜形成过程中algD基因表达水平的影响。结果:对PA17用1倍和10倍MIC的亚胺培南/西司他丁干预时,第3天algD表达较对照组分别增加1.5和3.4倍,生物膜形态无明显变化;第6天algD表达分别增加0.38和0.78倍,没有形成生物膜。对PAO1用1倍和10倍MIC的亚胺培南/西司他丁干预时,第24小时algD表达较对照组分别增加2.5和3.5倍,生物膜形态无明显变化;第3天algD分别增加0.7和2.1倍,1倍MIC干预组仍可看到少量微菌落存在,10倍MIC干预组仅见散在的细菌;第6天algD分别增加1.5和2.1倍,两个浓度组均只见数个细菌黏附于盖玻片。结论:亚胺培南/西司他丁对黏液型和非黏液型铜绿假单胞菌algD表达和生物膜形成的作用一致,虽可诱导algD表达上调从而使藻酸盐合成增加,但若从生物膜形成早期开始使用,仍可抑制和破坏生物膜形成。Objective：To study the effect of imipenem/cilastatin sodium on Pseudomonas aeruginosa （PA）17 and PAO1 in respect of biofilm formation and alginate biosynthesis. Methods： Tube fold dilution was used to detect the minimum inhibitory concentration （MIC） of imipenem/cilastatin on PAl7 and PAO1. Modified plate culture method was used to establish the biofilm model in vitro. The effect of 1×MIC and 10×MIC imipenem/cilastatin on PAl7 and PAO1 biofilm formation was observed by scanning electron microscope and semi quantitative RT-PCR was used to determine the effect on the gene expression of algD. Results： When PAl7 biofilm was exposed to 1×MIC and 10×MIC imipenem, algD expression level increased 1.5 fold and 3.4 fold respectively compared to the controls on the 3rd day, the biofilm structure change was not significant; the algD expression level increased 0.38 fold and 0. 78 fold respectively on the 6th day, and without biofilm formation. When PAO1 biofilm was exposed to 1×MIC and 10×MIC imipenem, the algD expression level increased 2.5 fold and 3.5 fold respectively compared to the controls at 24h, the morphology of biofilm showed no marked change; the algD expression level was increased 0.7 fold and 2.1 fold respectively on the 3rd day, there were a few microcolonies in 1×MIC group and only few scattered germs in 10×MIC group; the algD expression level was induced to 1.5 fold and 2.1 fold respectively on the 6th day, and without biofilm formation. Conclusion： The effect of imipenem/cilastatin sodium on biofilm formation and alginate biosynthesis of mucoid and nonmucoid Pseudomonas aeruginosa are the same. Although imipenem/cilastatin sodium can induce the expression of algD, it still can inhibit the Pseudomonas aeruginosa biofilm formation if it used in the early stage of biofilm formation.

关 键 词：铜绿假单胞菌 生物膜 藻酸盐 亚胺培南/西司他丁 

分 类 号：R515[医药卫生—内科学]