抗菌药物联合中药单体对泛耐药鲍曼不动杆菌生物被膜的影响

摘要：目的 研究4种抗菌药物(亚胺培南、美罗培南、多黏菌素B和替加环素) 和3种中药单体(黄芩苷、盐酸小檗碱和槲皮素二水物)对泛耐药鲍曼不动杆菌(XDRAB)生物被膜形成能力的影响，并探讨以上组合用药对XDRAB的协同抗生物被膜效应。方法 收集2018-2019年成都医学院第一附属医院的不同科室的临床标本分离的9株XDRAB，采用比浊法测定4种抗菌药物、3种中药单体在1 MIC、1/2 MIC、1/4 MIC、1/8 MIC、1/16 MIC对9株XDRAB 24 h内的生长情况；结晶紫棋盘染色法检测工作浓度下亚胺培南、美罗培南、替加环素、多黏菌素B、黄芩苷、盐酸小檗碱和槲皮素二水物单用对生物被膜的抑制作用和组合联用的协同抗生物被膜效应。结果 亚胺培南(4 μg/mL)、美罗培南(2 μg/mL)、多黏菌素B(0.25 μg/mL)、替加环素(0.0625 μg/mL)、黄芩苷(128 μg/mL)、盐酸小檗碱(32 μg/mL)、槲皮素二水物(64 μg/mL)的浓度时24 h内不抑制细菌的生长，以此浓度为基础倍比稀释，设置高、中、低三个浓度梯度；与模型对照组相比，4种抗菌药物、3种中药单体、在高浓度下单用、联用均可以明显抑制XDRAB生物被膜的形成(P＜0.05)，联合用药效果均优于单独用药(P＜0.05)，表现为不同程度的协同抗生物被膜作用，其中替加环素与黄芩苷的9个浓度组合均能够显著抑制XDRAB生物被膜的形成，0.015625 μg/mL替加环素与32 μg/mL黄芩苷与的浓度组合下，药物剂量最小。结论 4种抗菌药物亚胺培南、美罗培南、多黏菌素B、替加环素，3种中药单体黄芩苷、槲皮素二水物、盐酸小檗碱单用及联用时对XDRAB的生物被膜形成均具有不同程度的抑制作用，联合用药均表现为不同程度的协同抗生物被膜效应。

In vitro activity of antibacterial agents in combination with traditional Chinese medicine monomers on the biofilm of extensively drug-resistant Acinetobacter baumannii

Abstract Objective To research the activity of four antibacterial agents (imipenem, meropenem, polymyxin B, and tigecycline) and three kinds of traditional Chinese medicine monomers (baicalin, berberine hydrochloride, and quercetin dihydrate) on the biofilm formation ability of extensively drug-resistant Acinetobacter baumanii (XDRAB), and to investigate the synergistic anti-biofilm activity of the combinations. Methods Nine extensively drug-resistantAcinetobacter baumanii isolates were collected between 2018 to 2019 from the First Affiliated Hospital of Chengdu Medical College. The turbidimetric method was used to determine the curve growth of nine XDRAB at 1 MIC, 1/2 MIC, 1/4 MIC, 1/8 MIC, 1/16 MIC for four antibacterial agents and three kinds of traditional Chinese medicine monomers within 24 h. The crystal violet checkerboard method was used to investigate the inhibitory activity of imipenem, meropenem, tigecycline, polymyxin B, baicalin, berberine hydrochloride, and quercetin dihydrate on biofilms at working concentrations and to research the synergistic anti-biofilm activity of combinations of the above mentioned drugs. Results Imipenem, meropenem, polymyxin B, tigecycline, baicalin, berberine hydrochloride, and quercetin dihydrate at 4, 2, 0.25, 0.0625, 128, 32, and 64 μg/mL respectively did not inhibit the growth of bacteria within 24 h. Using this concentration as the basis for multiple dilutions, three concentration gradients of high, medium, and low were set. Compared with the model control group, single use or combined use at high concentrations of the four antibacterial agents and three kinds of traditional Chinese medicine monomers could significantly inhibit the formation of XDRAB biofilm (P<0.05), and the activity of combined agents was better than that of single use (P<0.05), showing different degrees of synergistic anti-biofilm activities. Among them, the nine combinations of tigecycline and baicalin could significantly inhibit the formation of XDRAB biofilms. The combination of 0.015625 μg/mL tigecycline and 32 μg/mL baicalin had the smallest dose. Conclusion Four antibacterial agents (imipenem, meropenem, polymyxin B, and tigecycline) and three kinds of traditional Chinese medicine monomers (baicalin, berberine hydrochloride, and quercetin dihydrate) had different degrees of inhibition on the formation of XDRAB biofilms when used alone and in combination, and the combinations showed different degrees of synergistic anti-biofilm