中文摘要：

目的建立铜绿假单胞菌生物膜（biofilm，BF）模型，探讨氨溴索对粘液型铜绿假单胞菌成熟BF的结构影响。方法平板培养法培养铜绿假单胞菌7d，得到成熟BF，用扫描电镜（scanning electron microscope，SEM）观察氨溴索对BF形态结构的影响；激光共聚焦显微镜（confocal laser scanning microscopy，CLSM）结合BF图像结构分析软件（image structure analyer，ISA）对BF结构参数分析；荧光显微镜定性观察氨溴索对BF内活菌的作用，并测定氨溴索单独作用以及与环丙沙星联用后BF内活菌的荧光强度。结果氨溴索作用后电镜观察可见BF被破坏，基质样物变稀疏，仅见少量散在细菌。ISA软件定量分析显示：2mg／ml氨溴索作用后，BF厚度、平均扩散距离（average diffusion distance，ADD）和结构熵（textual entropy，TE）均减少（P〈0．05）；区域孔率（areal porosity，AP）增加（P〈0．05）。0．75mg／ml氨溴索干预后，有同样趋势，但效应不如高浓度明显。用荧光显微镜定性观察，当氨溴索浓度大于0．49mg／ml时，BF内活菌数减少。同时，氨溴索与环丙沙星存在协同作用（F=15．1，P〈0．05），且随着氨溴索浓度升高，协同作用增强。结论氨溴索可影响铜绿假单胞菌成熟BF形态结构，从而增强抗生素的杀菌活性。

英文摘要：

Objective To observe the effect of ambroxol on the structure of Pseudomonas aeruginosa biofilm （BF） in an established biofilm model in vitro. Method Mature BF was formed after 7 days＇ culture of Pseudomonas aeruginosa on plate. The BF structure was observed by scanning electron microscope （SEM）. The parameters of BF structure were analyzed though pictures from confocal laser scanning microscopy （CLSM） with Image Structure Analyzer （ISA） software. Fluorescence intensity of the viable bacteria in BF treated by ambroxol alone and together with ciprofloxacin were determined with fluorescence microscopy. Results After the treatment of ambroxol, the BF was destroyed and the matrix outsides became thin and abnormal compared with the control by SEM, and small amount of bacteria dispersed. The results of ISA showed that with the treatment of 2 mg/ml ambroxol, BF was decreased in thickness, average diffusion distance （ADD） and textual entropy （TE） were decreased in comparison with the group without ambroxol treatment （ P 〈 0.05 ）, however areal porosity （AP） was increased （ P 〈 0.05 ）. Whereas group treated by 0.75 mg/ml ambroxol though not so the same tendency of these parameters was seen in the obvious. Fluorescence microscopy indicated the viable bacteria in BF was reduced significantly （ F = 18, P 〈 0.05 ） when ambroxol concentration was higher than 0.49 mg/ml, and ambroxol showed synergistic effect when combined with ciprofloxacin （F = 15.1 ,P 〈 0.05 ） in a dose-dependent manner. Conclusion Ambroxol can affect the BF structure of Pseudomonas aeruginosa, and thus enhance the bactericidal ability of antibiotics.