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目的探索鱼腥草素钠(SH)联合乙二胺四乙酸二钠(EDTA-Na2)对生物被膜菌的影响,寻找潜在的抗生素替代品。方法通过微量稀释法以及棋盘法,测定SH和EDTA-Na2对铜绿假单胞菌(Pseudomonas aeruginosa,PA)、金黄色葡萄球菌(Staphylococcus aureus,SA)、白色念珠菌(Candida albicans,CA)的最小抑菌浓度(MIC)和最小膜清除浓度(MBEC)以及两药协同点;通过平板稀释涂布法测定3种菌在协同点的生存曲线;扫描电子显微镜法(SEM)观察PA、SA和CA生物被膜的形态。结果 SH对PA、SA和CA的MIC分别为2.048、0.064、0.064 mg/m L,MBEC分别为2.048、0.256、0.512mg/m L;EDTA-Na2对PA、SA和CA的MIC分别为3.75、0.938、0.117 mg/m L,MBEC分别为15、3.75、30 mg/m L;SH联合EDTA-Na2(SH/EDTA-Na2)对PA、SA和CA的MIC(mg/m L)分别为0.256/0.938、0.008/0.233、0.008/0.029,MBEC(mg/m L)分别为0.512/3.722、0.032/0.469、0.064/0.938;SEM观察到两药联合对细菌的生长抑制和生物被膜的清除有明显作用,载体上的细菌显著减少,未见或仅有少量的生物被膜结构,与空白对照组比较差异明显。结论 SH与EDTA-Na2具有协同作用,二者联合能够显著增强SH对PA、SA和CA的生长抑制和生物被膜的清除作用。
Objective To explore the effect of sodium houttuyfonate (SH) and EDTA-Na2 on the biofilm and to find potential antibiotic alternatives. Methods The minimal inhibitory concentrations of SH and EDTA-Na2 against Pseudomonas aeruginosa (PA), Staphylococcus aureus (SA) and Candida albicans (CA) were determined by microdilution and checkerboard methods (MIC) and minimum membrane clearance concentration (MBEC) as well as the coordination point between the two drugs. The survival curves of the three bacteria in the synergy point were determined by plate dilution coating method. The scanning electron microscopy (SEM) Biofilm morphology. Results The MICs of SH, PA, SA and CA were 2.048, 0.064 and 0.064 mg / m L, respectively, and MBECs were 2.048,0.256 and 0.512 mg / m L, respectively. The MIC of EDTA, 0.938,0.117 mg / m L and MBEC were 15,3.75 and 30 mg / m L respectively. The MIC (mg / m L) of SH combined with EDTA-Na2 (SH / EDTA-Na2) /0.938,0.008/0.233,0.008/0.029, MBEC (mg / m L) were 0.512 / 3.722,0.032 / 0.469,0.064 / 0.938; SEM observation of the growth inhibition of the combination of bacteria and biofilm clear Role of the carrier significantly reduced bacteria, no or only a small amount of biological envelope structure, compared with the blank control group significant differences. Conclusion SH and EDTA-Na2 have a synergistic effect. The combination of SH and SH can significantly enhance the growth inhibition of PA, SA and CA and the scavenging effect of biofilm.