红霉素、氨溴索与环丙沙星雾化吸入对实验大鼠铜绿假单胞菌生物膜的影响

The intervention in endotracheal tube-associated biofilms and respiratory infections of Pseudomonas aeruginosa by using inhalation of Erythromycin and Ambroxol in combination with Ciprofloxacin respectively in rat model

目的研究红霉素和氨溴索分别联合环丙沙星雾化吸入对铜绿假单胞菌成熟生物膜的干预效果。方法平板法培养成熟铜绿假单胞菌生物膜;微量肉汤稀释法测量红霉素和环丙沙星的最低抑菌浓度;制作气管插管铜绿假单胞菌生物膜感染模型;平板计数法计算红霉素、氨溴索分别联合环丙沙星对生物膜菌落数的影响;日本岛津紫外-可见光分光光度计UV1700测铜绿假单胞菌菌液的A值;石蜡切片HE染色定性观察肺组织的炎症情况;扫描电镜定性观察各处理组的生物膜结构变化。结果各处理组干预7 d后肺组织细菌菌落计数(×104CFU/m l):干预组分为:生理盐水对照,氨溴索,红霉素,红霉素联合环丙沙星,氨溴索联合环丙沙星,各组分别为139.250±42.0162、101.625±40.4190、109.625±33.4747、57.750±37.8295和22.250±17.3184,前3组与后2组对比差异均有显著性(P<0.05),前3组之间对比差异没有显著性(P>0.05),后2组对比差异有显著性(P<0.05)。导管生物膜细菌菌落计数(×104CFU/m l):5组分别为170.000±48.3263、127.625±39.0163、133.500±33.6876、70.375±35.7768和38.125±19.1045,结论和肺组织菌落计数是一致的。导管生物膜电镜观察:第1组导管内表面均有较厚基质覆盖,2、3组减少不明显,而联合用药组导管内表面生物膜明显减少,其中第5组效果更好。结论氨溴索与红霉素分别联合环丙沙星雾化吸入在控制导管生物膜和呼吸系统相关感染均具有显著效果,其中氨溴索联合环丙沙星疗效更好。

Objective To research the effect of inhalation of Erythromycin and Ambroxol combined with Ciprofloxacin respectively against respiratory infection with endotracheal tube（ETT）-associated biofilm of Pseudomonas aeruginosa.Method Plate culture method was used to establish mature biofilm and tube dilution method to measure the Minimum Inhibitory Concentration（MIC） of Erythromycin and Ciprofloxacin.Rat model of Pseudomonas aeruginosa biofilm infection was established.The biofilm was treated by Erythromycin and Ambroxol combined with Ciprofloxac respectively.The number of the bacteria within biofilm was measured by agar plate.The visible light spectrophotometer was used to measure the optical density（OD） value of Pseudomonas aeruginosa.The inflammation of the lung was observed through paraffin section,hematoxylin and eosin stain（HE） stain.The variation of each group＇s biofilm was observed.Result After 7 days of intervention,the bacterial colony count（BCC） of lung tissues（×104CFU/ml） from each group（group1=control,group2=ambroxol,group3=erythromycin,group4=erythromycin＋ciprofloxacin,group5= ambroxol＋ciprofloxacin）,respectively,was 139.250±42.0162,101.625±40.4190,109.625±33.4747,57.750±37.8295 and 22.25±17.31840.There were significantly difference between the first three groups（group1,2 and 3） and the others respectively（P〈0.05）;There also was significantly difference between group4 and group5（P〈0.05）,while the differences between group1,group2 and group3 were not significant（P〉0.05）.BCC of ETT biofilm（×104CFU/ml） from each group was,in turn,170.000±48.3263,127.625±39.0163,133.500±33.6876,70.375±35.7768 and 38.125±19.1045.Scanning Electron Microscopy（SEM） results showed that the matrix of the ETT surface in group1 was the thickest,followed by those in group2 and group3,while those in group4 and group5 were much less,with that of group5 being the least.Con clusion The combination of Ambroxol with Ciprofloxac was signifcantly effective on ETT-associated biofilm of P. aeruginosa and relevant respiratory infection,which was better than the combination of Erythromycin with Ciproflox acin.