Efficacy of Calcium-EDTA as an Inhibitor for Metallo-β-Lactamase in a Mouse Model of Pseudomonas aeruginosa Pneumonia
Efficacy of Calcium-EDTA as an Inhibitor for Metallo-β-Lactamase in a Mouse Model of Pseudomonas aeruginosa Pneumonia
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DOI：
10.1128/aac.00511-10
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发表时间：
2010-11-01
影响因子：
4.9
通讯作者：
Yamaguchi, Keizo
中科院分区：
文献类型：
--
作者：
Aoki, Nobumasa;Ishii, Yoshikazu;Yamaguchi, Keizo
在这项研究中，我们在铜绿假单胞菌肺炎小鼠模型中评估了钙- edta （Ca-EDTA）作为细菌金属酶（如金属β -内酰胺酶（MBL）和其他蛋白酶）抑制剂的功效。同时存在的Ca-EDTA （32 μ g/ml）降低了所有产mbl铜绿假单胞菌（IMP-1、-2、-7、-10和VIM-2）的亚胺培烯（IPM）的mic，但对非产mbl菌株没有影响。在肺炎模型中，小鼠鼻内感染产生mbl的铜绿假单胞菌，然后保持在高氧条件下，以模拟呼吸机相关性肺炎。通过鼻内和皮下给药，与单独给药相比，Ca-EDTA显著增强了IPM的生存益处。Ca-EDTA联合治疗可显著降低肺部细菌负荷（P < 0.05）。此外，通过纯化的mp -1酶证实了Ca-EDTA对MBL活性的抑制作用，其50%抑制浓度（IC50）为55 +/- 8.2 μ m。最后，通过培养上清诱导的小鼠上皮细胞损伤和急性肺损伤，证实了Ca-EDTA的保护作用。这些数据表明，Ca-EDTA不仅可以阻断MBLs，还可以中和铜绿假单胞菌感染中的组织损伤金属蛋白酶，具有治疗潜力。
In this study, we have evaluated the efficacy of calcium-EDTA (Ca-EDTA) as an inhibitor of bacterial metalloenzymes, such as metallo-beta-lactamase (MBL) and other proteases, in a mouse model of Pseudomonas aeruginosa pneumonia. The simultaneous presence of Ca-EDTA (32 mu g/ml) reduced the MICs of imipenem (IPM) in all MBL-producing P. aeruginosa isolates (IMP-1, -2, -7, and -10 and VIM-2) but not non-MBL-producing strains. In the pneumonia model, mice were intranasally infected with MBL-producing P. aeruginosa and then kept under conditions of hyperoxia to mimic ventilator-associated pneumonia. With both intranasal and subcutaneous administrations, Ca-EDTA significantly potentiated survival benefits of IPM compared to those of IPM alone. Ca-EDTA combination therapy induced a significant reduction of the bacterial burden in the lungs (P < 0.05). Furthermore, the inhibition activity of Ca-EDTA against MBL activity was confirmed by using the purified IMP-1 enzyme, which was characterized by a 50% inhibitory concentration (IC50) of 55 +/- 8.2 mu M. Finally, the protective effects of Ca-EDTA were demonstrated by culture supernatant-induced epithelial cell damage and acute lung injury in mice. These data suggest the therapeutic potential of Ca-EDTA not only by the blocking of MBLs but also by neutralizing tissue-damaging metalloproteases in P. aeruginosa infections.