黄芩苷联合头孢哌酮/舒巴坦对铜绿假单胞菌生物膜破坏作用的体外研究

摘要

Objective To observe the destructive effect of baicalin on the biofilm of pseudomonas aeruginose and the sterilizing effect on the combination with sulbactam-cefperazone. Methods PAO1 was selected for biofilm formation in vitro. The minimum in inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) of baicalin and sulbactam-cefperazone were measured by doubling dilution;viable bacterial counts in biofilm were determined by serial dilution;the surface morphology of biofilm was observed by fluorescence microscope. Results When the biofilm being reacted with drugs for 12 hours,the viable bacterial counts showed the number of bacteria in biofilm of the baicalin group, sulbactam cefoperazone group, baicalin combined with cefoperazone and sulbactam was significantly less than that in the control group (P<0.01); baicalin combined with cefoperazone and sulbactam group was significantly less than that in the baicalin group and sulbactam-cefpera-zone group (P<0.01). While there was no statistical difference between baicalin group and sulbactam-cefperazone group (P>0.05). Fluorescence microscope revealed that the biofilm formed in the baicalin group which was almost viable cells (green fluo-rescence) was significantly less than that in the control group,the same as the sulbactam-cefperazone group whose green fluores-cence was more than the red one. While baicalin combined with sulbactam-cefperazone group were almost dead, which was al-most the red fluorescence. Conclusion Baicalin can destroy the formed pseudomonas aeruginose biofilm and significantly en-hance bactericidal effect on pseudomonas aeruginose biofilm when used combined with cefoperazone and sulbactam.%目的：研究黄芩苷对铜绿假单胞菌生物膜的破坏作用以及与头孢哌酮/舒巴坦联合应用的杀菌效果。方法选取铜绿假单胞野生株PAO1建立体外生物膜模型，二倍稀释法测定黄芩苷以及头孢哌酮/舒巴坦的最低抑菌浓度（MIC）和最低杀菌浓度（MBC）；连续活菌稀释法测定生物膜内活菌计数；荧光显微镜观察生物膜表面形态。结果生物膜形成加入药物12h后，活菌计数结果显示，无论药物单独作用还是联合应用时与空白组相比差异均有统计学意义（P<0.01）；黄芩苷联合头孢哌酮/舒巴坦组与黄芩苷组、头孢哌酮/舒巴坦组比较，生物膜内活菌数差异均有统计学意义（P<0.01）；黄芩苷组与头孢哌酮/舒巴坦组比较，生物膜内活菌数差异无统计学意义（P＞0.05）。荧光显微镜观察发现，黄芩苷组较空白对照组所形成的生物膜有所减少且分布较稀薄，且以活菌为主（绿色信号）；头孢哌酮/舒巴坦组较空白对照组所形成的生物膜分布稀薄，活菌为主，有少量死菌（红色信号）；黄芩苷联合头孢哌酮/舒巴坦组可观察到少量细菌生物膜，且主要是分布散落的细菌，多以死菌为主。结论黄芩苷能破坏铜绿假单胞菌已形成的生物膜，与头孢哌酮/舒巴坦联合应用时能显著增强其对铜绿假单胞菌生物膜的杀菌作用。