氨溴索对成熟表皮葡萄球菌生物被膜结构的破坏作用和膜内菌的杀灭作用  

Effect of ambroxol on the structure of Staphylococcus epidermidis biofilm and bactericidal action

作　　者：郜向娜[1] 余加林[1] 吴玉华[1] 朱秀菊[1] 刘维勤[1] 

机构地区：[1]重庆医科大学附属儿童医院新生儿科儿童发育疾病研究省部共建教育部重点实验室,重庆400014

出　　处：《中国抗生素杂志》2011年第8期635-639,共5页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(编号:30772363;81070513);2008年教育部博士基金(编号:20070631001);重庆市科委自然基金(编号:2007BB5282);重庆医科大学2008年校级重点项目(编号:XBZD200808)

摘　　要：目的建立表皮葡萄球菌(S.epidermidis)生物被膜(biofilm,BF)体外模型,探讨氨溴索对表皮葡萄球菌成熟BF结构的破坏作用和对膜内菌的杀灭作用。方法平板法培养表皮葡萄球菌RP62A 24h,得到成熟BF,不同浓度氨溴索作用24h后,用扫描电镜(scanning electron microscope,SEM)观察BF的形态结构;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结构参数进行分析,利用四唑盐(XTT)减低法检测氨溴索对BF内细菌的杀灭作用。结果氨溴索处理以后,电镜观察可见1.875mg/mL氨溴索处理组BF结构被破坏,仅见少量散在细菌。ISA软件定量分析显示:1.875mg/mL氨溴索处理组和对照组的BF厚度分别为(27.95±5.93)μm和(9.76±1.29)μm,t=7.955,P<0.05;平均扩散距离(average diffusion distance,ADD)分别为2.173±0.219和1.625±0.092,t=5.640,P<0.05;结构熵(textual entropy,TE)分别为7.706±0.610和5.488±0.747,t=5.633,P<0.05;区域孔率(areal porosity,AP)分别为0.772±0.036和0.889±0.063,t=-4.322,P<0.05。XTT减低法显示氨溴索的浓度为1.875、3.7和7.5mg/mL时,A450的值分别为1.288±0.251、0.548±0.170和0.156±0.05,与对照组相比P<0.05,均有统计学意义。结论氨溴索可破坏表皮葡萄球菌BF的形态结构,随着氨溴索浓度的增加,杀灭BF内细菌的能力增加。Objective To investigate the effect of ambroxol on Staphylococcus epidermidis biofilm （BF） in an established biofilm model in vitro. Methods The strain RP62A of Staphylococcus epidermidis was incubated for 24h to form BF on slide. After treated by ambroxol of different concentrations for 24h, the BF structure was observed by scanning electron microscope （SEM）, the parameters of BF structure were analyzed through pictures from Confocal laser scanning microscopy （CLSM） with image structure analyer （ISA） software; the metabolic activity of bacteria in biofilm was measured by tetrazolium salt （XTT） reduction assay. Results After the treatment of 1.875mg/mL ambroxol, the BF was destroyed and the matrix outsides became thin and sparseness compared with the control by SEM, and small amount of bacteria scattered. BF thickness of the 1.875mg/mL ambroxol treatment group and control group were （27.95±5.93）gm and （9.76±1.29）gm, respectively., t=7.955, P〈0.05, areal porosity（AP） were 0.7724±0.036 and 0.889±0.063 respectively, t=-4.322, P〈0.05; average diffusion distance（ADD） were 2.173±0.219 and 1.625±0.092 respectively, t=5.640, P〈0.05,and textual entropy（TE） were 7.706±0.610 and 5.488±0.747 respectively, t=5.633, P〈0.05; The effect of ambroxol 0.75mg/mL has the same tendency as ambroxol 1.875 mg/mL group, but not so obvious. After the administration by ambroxol of different concentration （1.875, 3.7 and 7.5mg/mL）, the A450 value were 1.288±0.251, 0.548±0.170 and 0.156±0.051 respectively. There were significant differences between each group and the control group （P〈0.05）. Conelusion Ambroxol can destroy the BF structure of Staphylococcus epidermidis. With increasing concentration of ambroxol, the bactericidal activity increased.

关 键 词：表皮葡萄球菌 生物被膜 氨溴索 XTT 

分 类 号：R378.11[医药卫生—病原生物学]