【精品推荐】盐酸氨溴索联合环丙沙星抗铜绿假单胞菌成熟生物膜作用研究药学论文_医药学论文_25055.doc
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1、论文范文题目:盐酸氨溴索联合环丙沙星抗铜绿假单胞菌成熟生物膜作用研究药学论文_医药学论文编辑:小小作者:李芳 余加林 杨华 李禄全 陈波曼【摘要】 目的 构建铜绿假单胞菌发光菌株,建立铜绿假单胞菌生物膜(biofilm,BF)模型,研究盐酸氨溴索对铜绿假单胞菌成熟BF的影响及与环丙沙星合用是否具有协同杀菌作用。方法 将携带绿色荧光蛋白(green fluorescent protein,GFP)的质粒转染铜绿假单胞菌,平板培养法培养铜绿假单胞菌BF,建立铜绿假单胞菌BF体外模型,经盐酸氨溴索作用后,扫描电镜(scanning electron microscope,SEM)观察盐酸氨溴索对BF
2、形态结构的影响,荧光显微镜定性观察盐酸氨溴索对BF内活菌的作用,通过发光菌株荧光强度定量分析盐酸氨溴索单独作用以及与环丙沙星联用后BF内活菌量。结果 盐酸氨溴索作用后电镜观察可见黏液样物变稀疏,不规则。盐酸氨溴索浓度大于0.49mg/mlBF内活菌数减少(F=18,P0.05);盐酸氨溴索与环丙沙星存在协同作用(F=15.1,P0.05),且随着盐酸氨溴索浓度升高协同作用增强。结论 盐酸氨溴索破坏铜绿假单胞菌成熟BF形态结构,减少其内活菌数;盐酸氨溴索与环丙沙星存在协同作用,增强其杀菌能力。 【关键词】 盐酸氨溴索; 环丙沙星; 铜绿假单胞菌; 生物膜ABSTRACT Objective To
3、 establish a biofilm model of Pseudomonas aeruginosa in vitro and observe the effect of ambroxol on biofilm of Pseudomonas aeruginosa, and study the synergistic effect with ciprofloxacin on tested specimens. Method Plasmids carried with green fluorescent protein (GFP) was tansfected to the strain of
4、 Pseudomonas aeruginosa. Plate culture method was used to establish biofilm model of Pseudomonas aeruginosa in vitro. After ambroxol was added to the biofilm, the appearance of the biofilm was detected by scanning electron microscope (SEM), and the viable counts of bacterial in biofilm after treated
5、 by ambroxol alone and together with ciprofloxacin were determined by bio-fluorescence method. Result After adding ambroxol, the biofilm was thin and abnormal compared with the control by SEM detection. Further detection with fluorescence microscope showed the viable bacteria in biofilm was reduced
6、significantly (F=18, P0.05) when its concentration was higher than 0.49 mg/ml, destroyed furthermore, there was the synergistic effect between ambroxol and ciprofloxacin (F=15.1, P0.05). The effect enhanced with the increasing concentration of ambroxol. Conclusion Ambroxol destroyed furthermore the
7、biofilm shape of Pseudomonas aeruginosa, restrained its growth, and had the synergistic effect on bactericidal activity with ciprofloxacin.KEY WORDS Ambroxol; Ciprofloxacin; Pseudomonas aeruginosa; Biofilm生物膜(biofilm,BF)是细菌黏附在物体表面上形成的一种具有高度结构性的膜状复合物,其主要成分是细菌分泌的胞外基质和细菌菌体1。BF可使细菌具有抵御宿主免疫系统和抗菌药物杀伤作用,产生
8、对抗菌药物的高度耐药性(可达到浮游状态的101000倍),导致感染难以治愈2。铜绿假单胞菌是常见细菌生物膜相关感染病原菌之一,铜绿假单胞菌生物膜的形成是呼吸机相关性感染难以控制的重要原因。作者曾研究氨溴索可显著促进环丙沙星对细菌生物膜的透过3,本研究发现盐酸氨溴索对铜绿假单胞菌BF有抑制作用,且与环丙沙星联用后具有协同作用,增强杀菌活性,现报道如下。1 材料和方法1.1 材料菌株PAO1(购自四川大学微生物实验室),Luria-Bertani培养基(LB,上海升博生物技术公司),盐酸氨溴索标准品(德国Boehringer Ingelheim);环丙沙星标准品(重庆市药品检定所),比浊仪(德国S
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