Browse > Article
http://dx.doi.org/10.5483/BMBRep.2014.47.4.040

siRNA-mediated gene silencing of MexB from the MexA-MexB-OprM efflux pump in Pseudomonas aeruginosa  

Gong, Feng-Yun (Department of Infectious Diseases, Wuhan Pu-ai Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Zhang, Ding-Yu (Department of Anesthesiology, Wuhan Pu-ai Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Zhang, Jiang-Guo (Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Wang, Li-Li (Department of Respiratory Medicine, Central Hospital of Qingdao)
Zhan, Wei-Li (Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Qi, Jun-Ying (Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Song, Jian-Xin (Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology)
Publication Information
BMB Reports / v.47, no.4, 2014 , pp. 203-208 More about this Journal
Abstract
To gain insights into the effect of MexB gene under the short interfering RNA (siRNA), we synthesized 21 bp siRNA duplexes against the MexB gene. RT-PCR was performed to determine whether the siRNA inhibited the expression of MexB mRNA. Changes in antibiotic susceptibility in response to siRNA were measured by the E-test method. The efficacy of siRNAs was determined in a murine model of chronic P. aeruginosa lung infection. MexB-siRNAs inhibited both mRNA expression and the activity of P. aeruginosa in vitro. In vivo, siRNA was effective in reducing the bacterial load in the model of chronic lung infection and the P. aeruginosa-induced pathological changes. MexB-siRNA treatment enhanced the production of inflammatory cytokines in the early infection stage (P < 0.05). Our results suggest that targeting of MexB with siRNA appears to be a novel strategy for treating P. aeruginosa infections.
Keywords
Antibiotic sensitivity; Efflux pump; MexB; Pseudomonas aeruginosa; siRNA;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Yanagihara, K., Tashiro, M. and Fukuda, Y. (2006) Effects of short interfering RNA against methicillin-resistant Staphylococcus aureus coagulase in vitro and in vivo. J. Antimicrob. Chemother. 57, 122-126.   DOI
2 Coban, A. Y., Ekinci, B. and Durpinar, B. (2004) A multidrug efflux pump inhibitor reduces fluoroquinolone resistance in Pseudomonas aerugnosa isolates. Chemotherap. 50, 22-26.
3 Hong, W., Song, Z., Givskov, M., Doring, G., Worlitzsch, D., Mathee, K., Rygaard, J. and Hoiby, N. (2001) Pseudomonas aeruginosa mutations in lasI and rhlI quorum sensing systems result in milder chronic lung infection. Microbiology 147, 1105-1113.   DOI
4 Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E. and Mello, C. C. (1998) Potent and genetic in-terference by double stranded RNA in caenorhabditis elegans. Nature 391, 806-811.   DOI   ScienceOn
5 Takeuchi, K., Kiefer, P., Reimmann, C., Keel, C., Dubuis, C., Rolli, J., Vorholt, J. A. and Haas, D. (2009) Small RNA-dependent Expression of Secondary Metabolism Is Controlled by Krebs Cycle Function in Pseudomonas fluorescens. J. Biol. Chem. 284, 34976-34985.   DOI   ScienceOn
6 Aira, H. (2007) Mechanism of RNA silencing by Hfq-binding small RNAs. Curr. Opin. Microbiol. 10, 134-139.   DOI   ScienceOn
7 Sonnleitner, E. and Haas, D. (2011) Small RNAs as regulators of primary and secondary metabolism in Pseudomonas species. Appl. Microbiol. Biotechnol. 91, 63-79.   DOI
8 Waqner, J. G. and Roth, R. A. (2000) Neutrophil migration mechanisms,with an emphasis on the pulmonary vasculature. Pharmacol. Rev. 52, 349-374.
9 Moser, C., Jensen, P. O., Kobayashi, O., Hougen, H. P., Song, Z., Rygaard, J. and Kharazmi, A. H. by N. (2002) Improved outcome of chronic Pseudomonas aeruginosa lung infection is associated with induction of a Th1- dominated cytokine response. Clin. Exp. Immunol. 127, 206-213.   DOI
10 Reiniger, Nina., Lee, M. M., Coleman, F. T., Ray, C., Golan, D. E. and Pier, G. B. (2007) Resisitance to Pseudomonas aeruginosa chronic lung infection requires cystic fibrosis transmembrane conductance regulatormodulated interleukin-1 (IL-1) release and signaling through the IL-1 receptor. Infect. Immun. 75, 1598-1608.   DOI   ScienceOn
11 Li, X. Z., Zhang, L. and Poole, K. (2000) Interplay between the MexA-MexB-OprM multidrug efflux system and the outer membrane barrier in the multiple antibiotic resistance of Pseudomonas aeruginosa. Antimicrob. Chemother. 45, 433-436.   DOI   ScienceOn
12 Sanchez, P., Rojo, F. and Martinez, J. L. (2002) Transcriptional regulation of MexR, the repressor of Pseudomonas aeruginosa MexAB-OprM multidrug efflux pump. FEMS. Microbiol. Lett. 207, 63-68.   DOI   ScienceOn
13 Strieter, R. M., Standiford, T. J. and Huffnagle, G. B. (1996) ''The good, the bad and the ugly'', The role of chemokines in models of human disease. J. Immunol. 156, 3583-3586.
14 Koedel, U., Frankenberg, T., Kirschnek, S., Obermaier, B., Hacker, H., Paul, R. and Hacker, G. (2009) Apoptosis Is Essential for Neutrophil Functional Shutdown and Determines Tissue Damage in Experimental Pneumococcal Meningitis. PloS Pathog. 5, e1000461.   DOI   ScienceOn
15 Perez-Martinez, I. and Haas, D. (2011) Azithromycin Inhibits Expression of the GacA-Dependent Small RNAs RsmY and RsmZ in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 55, 3399-3405.   DOI   ScienceOn
16 Wang, Y. and Cela, E. (2010) Estrogen aggravates inflammation in Pseudomonas aeruginosa pneumonia in cystic fibrosis mice. Respir. Res. 11, 166.   DOI   ScienceOn
17 Araki, N., Yanagihara, K., Morinaga, Y., Yamada, K., Yamada, Y., Kohno, S. and Kamihira, S. (2011) In vivo efficacy of doripenem (DRPM) against Pseudomonas aeruginosa in murine chronic respiratory tract infection model. J. Infect. Chemother. 17, 318-321.   DOI
18 Macia, M. D., Borrell, N., Perez, J. L. and Oliver, A. (2004) Detection and Susceptibility Testing of Hypermutable Pseudomonas aeruginosa Strains with the Etest and Disk Diffusion. Antimicrob. Agents Chemother. 48, 2665-2672.   DOI   ScienceOn
19 Wu, C. M., Cao, J. L., Zheng, M. H., Ou, Y., Zhang, L., Zhu, X. Q. and Song, J. X. (2008) Effect and Mechanism of Andrographolide on the Recovery of Pseudomonas aeruginosa Susceptibility to Several Antibiotics. J. Int. Med. Res. 36, 178-186.   DOI
20 Strateva, T. and Yordanov, D. (2009) Pseudomonas aeruginosa-a phenomenon of bacterial resistance. J. Med. Microbiol. 58, 1133-1148.   DOI   ScienceOn
21 Srikumar, R., Kon, T., Gotoh, N. and Poole, K. (1998) Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain. Antimicrob. Agents. Chemother. 42, 65-71.
22 Liu, Y. S., Zhang, Y. Q. and Xiao, C. L. (2009) Gene-specific silencing induced by parallel complementary RNA in Pseudomonas aeruginosa. Biotechnol. Lett. 31, 1571-1575.   DOI
23 Srikumar, R., Kon, T., Gotoh, N. and Poole, K. (1998) Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain. Antimicrob. Agents Chemother. 42, 65-71.
24 Greenfield, T. J., Franch, T., Gerdes, K. and Weaver, K. E. (2001) Antisense RNA regulation of the par post-segregational killing system: structural analysis and mechanism of binding of the antisense RNA, RNAII and its target, RNAI. Mol. Microbiol. 42, 527-537.   DOI   ScienceOn
25 Mack, D., Siemssen, N. and Laufs, R. (1992) Parallel induction by glucose of adherence and a polysaccharide antigen specific for plastic-adherent Staphylococcus epidermidis: evidence for functional relation to intercellular adhesion. Infect. Immun. 60, 2048-2057.
26 Taniguchi, K., Ono, T. and Murakami, K. (2003) Novel Pseudomonas aeruginosa Gene That Suppresses Tolerance to Carbapenems. Antimicrob. Agents. Chemother. 47, 2997-3001.   DOI
27 Morinaga,Y., Yanagiharal, K. and Nakamura, S. (2008) In vivo efficacy and pharmacokinetics of tomopenem (CS-023), a novel carbapenem, against Pseudomonas aeruginosa in a murine chronic respiratory tract infection model. J. Antimicrob. Chemother. 62, 1326-1331.   DOI   ScienceOn
28 Kukavica-Ibrulj, I., Bragonzi, A., Paroni, M., Winstanley, C., Sanschagrin, F., O'Toole, G. A. and Levesque, R. C. (2008) In Vivo Growth of Pseudomonas aeruginosa Strains PAO1 and PA14 and the Hypervirulent Strain LESB58 in a Rat Model of Chronic Lung Infection. J. Bacteriol. 190, 2804-2813.   DOI   ScienceOn
29 Hazlett, L. D., Zucker, M. and Berk, R. S. (1992) Distribution and kinetics of the inflammatory cell response to ocular challenge with Pseudomonas aeruginosa in susceptible and resistant mice. Ophthalmic Res. 24, 32-39.
30 Poole, K. (2005) Efflux-mediated antimicrobial resistance. J. Antimicrob. Chemother. 56, 20-51.   DOI   ScienceOn