Gi Mia, Jeong Junhui, Lee Keehoon, Lee Kang-Mu, Toyofuku Masanori, Yong Dong Eun, Yoon Sang Sun, Choi Jae Young
Department of Otorhinolaryngology, Brain Korea 21 PLUS Project for Medical Science, Seoul, South Korea.
Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Seoul, South Korea.
Antimicrob Agents Chemother. 2014 Dec;58(12):7205-14. doi: 10.1128/AAC.03063-14. Epub 2014 Sep 22.
Pseudomonas aeruginosa, a Gram-negative bacterium of clinical significance, produces elastase as a predominant exoprotease. Here, we screened a library of chemical compounds currently used for human medication and identified diethylene triamine penta-acetic acid (DTPA, pentetic acid) as an agent that suppresses the production of elastase. Elastase activity found in the prototype P. aeruginosa strain PAO1 was significantly decreased when grown with a concentration as low as 20 μM DTPA. Supplementation with Zn(2+) or Mn(2+) ions restored the suppressive effect of DTPA, suggesting that the DTPA-mediated decrease in elastase activity is associated with ion-chelating activity. In DTPA-treated PAO1 cells, transcription of the elastase-encoding lasB gene and levels of the Pseudomonas quinolone signal (PQS), a molecule that mediates P. aeruginosa quorum sensing (QS), were significantly downregulated, reflecting the potential involvement of the PQS QS system in DTPA-mediated elastase suppression. Biofilm formation was also decreased by DTPA treatment. When A549 alveolar type II-like adenocarcinoma cells were infected with PAO1 cells in the presence of DTPA, A549 cell viability was substantially increased. Furthermore, the intranasal delivery of DTPA to PAO1-infected mice alleviated the pathogenic effects of PAO1 cells in the animals. Together, our results revealed a novel function for a known molecule that may help treat P. aeruginosa airway infection.
铜绿假单胞菌是一种具有临床意义的革兰氏阴性菌,它产生弹性蛋白酶作为主要的胞外蛋白酶。在此,我们筛选了一个目前用于人类药物治疗的化合物库,并确定二乙烯三胺五乙酸(DTPA,喷替酸)是一种抑制弹性蛋白酶产生的试剂。当在低至20μM DTPA的浓度下生长时,在原型铜绿假单胞菌菌株PAO1中发现的弹性蛋白酶活性显著降低。补充锌离子或锰离子可恢复DTPA的抑制作用,这表明DTPA介导的弹性蛋白酶活性降低与离子螯合活性有关。在经DTPA处理的PAO1细胞中,编码弹性蛋白酶的lasB基因的转录以及介导铜绿假单胞菌群体感应(QS)的分子铜绿假单胞菌喹诺酮信号(PQS)的水平均显著下调,这反映了PQS QS系统可能参与DTPA介导的弹性蛋白酶抑制作用。DTPA处理也降低了生物膜的形成。当在DTPA存在的情况下用PAO1细胞感染A549 II型肺泡样腺癌细胞时,A549细胞活力显著增加。此外,将DTPA经鼻递送至PAO1感染的小鼠可减轻PAO1细胞在动物体内的致病作用。总之,我们的结果揭示了一种已知分子的新功能,这可能有助于治疗铜绿假单胞菌气道感染。