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小柴胡汤及亚胺培南西司他丁对耐碳青霉烯类鲍曼不动杆菌的体外抑菌效果及生物膜清除作用的机制研究

Mechanism on antibacterial effect in vitro and biofilm eradication effect of Xiaochaihu Decoction and imipenem-cilastatin for carbapenem-resistant Acinetobacter baumannii

    摘要
  • 摘要:
    目的 分析小柴胡汤及亚胺培南西司他丁对耐碳青霉烯类鲍曼不动杆菌(CRAB)体外抑菌效果及生物膜清除作用的机制。
    方法 随机选取不同样本来源的60株CRAB。采用微量棋盘稀释法检测不同浓度组合的小柴胡汤联合亚胺培南西司他丁对60株CRAB的最低抑菌浓度(MIC), 并计算抑菌浓度指数(FIC)。采用酶标仪检测抗菌药物对生物膜的抑制作用; 采用实时荧光定量聚合酶链式反应(RT-PCR)检测耐药基因转录水平。
    结果 小柴胡汤和亚胺培南西司他丁对CRAB的MIC分别为32、16 μg/mL。小柴胡汤和亚胺培南西司他丁联合应用具有协同抑制CRAB生长的作用。与单独应用1/16 MIC亚胺培南西司他丁比较，加入1/16 MIC、1/4 MIC或1 MIC小柴胡汤联合1/16 MIC亚胺培南西司他丁后的A570值下降，差异有统计学意义(t=4.513、10.515、19.802, P < 0.05); 与单独应用1/16 MIC小柴胡汤比较，加入1/16 MIC、1/4 MIC或1 MIC亚胺培南西司他丁联合1/16 MIC小柴胡汤后的A570值下降，差异有统计学意义(t=3.109、5.657、15.461，P < 0.05)。与生长对照组相比，亚胺培南西司他丁组中adeJ和CarO呈低表达，联合组adeJ和CarO呈高表达，差异均有统计学意义(P < 0.05)。
    结论 小柴胡汤联合亚胺培南西司他丁具有协同抑制CRAB生长以及降低生物膜黏附性的作用，其机制可能是通过激活adeJ和CarO基因表达而实现的。

     

    Abstract:
    Objective To analyze the mechanism on antibacterial effect in vitro and biofilm eradication effect of Xiaochaihu Decoction and imipenem-cilastatin for carbapenem-resistant Acinetobacter baumannii (CRAB).
    Methods A total of 60 strains of CRAB isolated from different samples were randomly collected. The minimal inhibitory concentrations (MIC) of Xiaochaihu Decoction combined with imipenem-cilastatin in different concentration combinations for 60 CRAB strains were detected by micro-checkerboard dilution method, and fractional inhibitory concentration (FIC) was calculated. The inhibition effect of antimicrobial agents for biofilm was detected by microplate reader; real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was used to detect the transcription level of drug-resistant genes.
    Results MIC values of Xiaochaihu Decoction and imipenem-cilastatin for CRAB were 32 and 16 μg/mL, respectively. The combination of Xiaochaihu Decoction and imipenem-cilastatin was able to synergistically inhibit the growth of CRAB. Compared with 1/16 of MIC imipenem-cilastatin alone, the A570 value decreased after adding 1/16 MIC, 1/4 MIC or 1 of MIC of Xiaochaihu Decoction combined with 1/16 of MIC of imipenem-cilastatin, the difference was statistical significant (t=4.513, 10.515, 19.802, P < 0.05); compared with 1/16 of MIC of Xiaochaihu Decoction alone, the A570 value decreased after adding 1/16 of MIC, 1/4 of MIC or 1 of MIC of imipenem-cilastatin combined with 1/16 of MIC of Xiaochaihu Decoction, and the difference was statistically significant (t=3.109, 5.657, 15.461, P < 0.05). Compared with growth control group, expression levels of adeJ and CarO were significantly lower in the imipenem-cilastatin group, while their expression levels were significantly higher in the combination group(P < 0.05).
    Conclusion Xiaochaihu Decoction combined with imipenem-cilastatin can synergistically inhibit the growth of CRAB and reduce the adhesion of biofilm, and the mechanism may be achieved by activating the expression of adeJ and CarO genes.

     

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