Gnanadhas Divya Prakash, Elango Monalisha, Datey Akshay, Chakravortty Dipshikha
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India.
Department of Aerospace Engineering, Indian Institute of Science, Bangalore, India.
Sci Rep. 2015 Nov 2;5:16043. doi: 10.1038/srep16043.
Bacterial biofilms are associated with 80-90% of infections. Within the biofilm, bacteria are refractile to antibiotics, requiring concentrations >1,000 times the minimum inhibitory concentration. Proteins, carbohydrates and DNA are the major components of biofilm matrix. Pseudomonas aeruginosa (PA) biofilms, which are majorly associated with chronic lung infection, contain extracellular DNA (eDNA) as a major component. Herein, we report for the first time that L-Methionine (L-Met) at 0.5 μM inhibits Pseudomonas aeruginosa (PA) biofilm formation and disassembles established PA biofilm by inducing DNase expression. Four DNase genes (sbcB, endA, eddB and recJ) were highly up-regulated upon L-Met treatment along with increased DNase activity in the culture supernatant. Since eDNA plays a major role in establishing and maintaining the PA biofilm, DNase activity is effective in disrupting the biofilm. Upon treatment with L-Met, the otherwise recalcitrant PA biofilm now shows susceptibility to ciprofloxacin. This was reflected in vivo, in the murine chronic PA lung infection model. Mice treated with L-Met responded better to antibiotic treatment, leading to enhanced survival as compared to mice treated with ciprofloxacin alone. These results clearly demonstrate that L-Met can be used along with antibiotic as an effective therapeutic against chronic PA biofilm infection.
80%至90%的感染与细菌生物膜有关。在生物膜内,细菌对抗生素具有耐药性,所需浓度比最低抑菌浓度高1000倍以上。蛋白质、碳水化合物和DNA是生物膜基质的主要成分。主要与慢性肺部感染相关的铜绿假单胞菌(PA)生物膜含有细胞外DNA(eDNA)作为主要成分。在此,我们首次报告0.5μM的L-甲硫氨酸(L-Met)可抑制铜绿假单胞菌(PA)生物膜的形成,并通过诱导DNase表达来分解已形成的PA生物膜。在L-Met处理后,四个DNase基因(sbcB、endA、eddB和recJ)高度上调,同时培养上清液中的DNase活性增加。由于eDNA在建立和维持PA生物膜中起主要作用,DNase活性可有效破坏生物膜。用L-Met处理后,原本顽固的PA生物膜现在对环丙沙星敏感。这在小鼠慢性PA肺部感染模型的体内实验中得到了体现。与单独用环丙沙星治疗的小鼠相比,用L-Met治疗的小鼠对抗生素治疗反应更好,存活率更高。这些结果清楚地表明,L-Met可与抗生素一起用作治疗慢性PA生物膜感染的有效疗法。