The discovery of quorum sensing in Proteobacteria and its function in regulating virulence determinants makes it an attractive alternative towards attenuation of bacterial pathogens. In this study, crude extracts of Phyllanthus amarus Schumach. & Thonn, a traditional Chinese herb, were screened for their anti-quorum sensing properties through a series of bioassays. Only the methanolic extract of P. amarus exhibited anti-quorum sensing activity, whereby it interrupted the ability of Chromobacterium violaceum CVO26 to response towards exogenously supplied N-hexanoylhomoserine lactone and the extract reduced bioluminescence in E. coli [pSB401] and E. coli [pSB1075]. In addition to this, methanolic extract of P. amarus significantly inhibited selected quorum sensing-regulated virulence determinants of Pseudomonas aeruginosa PA01. Increasing concentrations of the methanolic extracts of P. amarus reduced swarming motility, pyocyanin production and P. aeruginosa PA01 lecA::lux expression. Our data suggest that P. amarus could be useful for attenuating pathogens and hence, more local traditional herbs should be screened for its anti-quorum sensing properties as their active compounds may serve as promising anti-pathogenic drugs.
References
[1]
Fuqua, C.; Parsek, M.R.; Greenberg, E.P. Regulation of gene expression by cell-to-cell communication: Acyl-homoserine lactone quorum sensing. Annu. Rev. Genet. 2001, 35, 439–468.
[2]
Kalia, V.C. Quorum sensing inhibitors: An overview. Biotechnol. Adv. 2013, 31, 224–245.
[3]
Pesci, E.C.; Pearson, J.P.; Seed, P.C.; Iglewski, B.H. Regulation of las and rhl quorum sensing in Pseudomonas aeruginosa. J. Bacteriol. 1997, 179, 3127–3132.
[4]
Gallagher, L.A.; Mcknight, S.L.; Kuznetsova, M.S.; Pesci, E.C.; Manoil, C. Functions required for extracellular quinolone signaling by Pseudomonas aeruginosa. J. Bacteriol. 2002, 184, 6472–6480.
[5]
Balasubramanian, D.; Schneper, L.; Kumari, H.; Mathee, K. A dynamic and intricate regulatory network determines Pseudomonas aeruginosa virulence. Nucleic Acids Res. 2013, 41, 1–20.
[6]
Winzer, K.; Falconer, C.; Garber, N.C.; Diggle, S.P.; Camara, M.; Williams, P. The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS. J. Bacteriol. 2000, 182, 6401–6411.
[7]
Lau, G.W.; Hassett, D.J.; Ran, H.; Kong, F. The role of pyocyanin in Pseudomonas aeruginosa infection. Trends Mol. Med. 2004, 10, 599–606.
[8]
Ochsner, U.A.; Koch, A.K.; Fiechter, A.; Reiser, J. Isolation and characterization of a regulatory gene affecting rhamnolipid biosurfactant synthesis in Pseudomonas aeruginosa. J. Bacteriol. 1994, 176, 2044–2054.
[9]
Brint, J.M.; Ohman, D.E. Synthesis of multiple exoproducts in Pseudomonas aeruginosa is under the control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer-responsive LuxR-LuxI family. J. Bacteriol. 1995, 177, 7155–7163.
Norizan, S.N.M.; Yin, W.F.; Chan, K.G. Caffeine as a potential quorum sensing inhibitor. Sensors 2013, 13, 5117–5129.
[13]
Tan, L.Y.; Yin, W.F.; Chan, K.G. Silencing quorum sensing through extracts of Melicope lunu-ankenda. Sensors 2012, 12, 4339–4351.
[14]
Krishnan, T.; Yin, W.-F.; Chan, K.-G. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa PAO1 by Ayurveda spice clove (Syzygium aromaticum) bud extract. Sensors 2012, 12, 4016–4030.
[15]
Taganna, J.C.; Quanico, J.P.; Perono, R.M.G.; Amor, E.C.; Rivera, W.L. Tannin-rich fraction from Terminalia catappa inhibits quorum sensing (QS) in Chromobacterium violaceum and the QS-controlled biofilm maturation and LasA staphylolytic activity in Pseudomonas aeruginosa. J. Ethnopharmacol. 2011, 134, 865–871.
[16]
Ali, H.; Houghton, P.J.; Soumyanath, A. alpha-Amylase inhibitory activity of some Malaysian plants used to treat diabetes; with particular reference to Phyllanthus amarus. J. Ethnopharmacol. 2006, 107, 449–455.
[17]
Naaz, F.; Javed, S.; Abdin, M.Z. Hepatoprotective effect of ethanolic extract of Phyllanthus amarus Schum. et Thonn. on aflatoxin B1-induced liver damage in mice. J. Ethnopharmacol. 2007, 113, 503–509.
[18]
Raphael, K.R.; Kuttan, R. Inhibition of experimental gastric lesion and inflammation by Phyllanthus amarus extract. J. Ethnopharmacol. 2003, 87, 193–197.
[19]
Kumar, K.B.H.; Kuttan, R. Chemoprotective activity of an extract of Phyllanthus amarus against cyclophosphamide induced toxicity in mice. Phytomedicine 2005, 12, 494–500.
[20]
McClean, K.H.; Winson, M.K.; Fish, L.; Taylor, A.; Chhabra, S.R.; Camara, M.; Daykin, M.; Lamb, J.H.; Swift, S.; Bycroft, B.W.; et al. Quorum sensing and Chromobacterium violaceum: Exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 1997, 143, 3703–3711.
[21]
Winson, M.K.; Swift, S.; Fish, L.; Throup, J.P.; J?rgensen, F.; Chhabra, S.R.; Bycroft, B.W.; Williams, P.; Stewart, G.S. Construction and analysis of luxCDABE-based plasmid sensors for investigating N-acyl homoserine lactone-mediated quorum sensing. FEMS Microbiol. Lett. 1998, 163, 185–192.
[22]
Hayouni, E.A.; Bouix, M.; Abedrabba, M.; Leveau, J.-Y.; Hamdi, M. Mechanism of action of Melaleuca armillaris [Sol. Ex Gaertu] Sm. essential oil on six LAB strains as assessed by multiparametric flow cytometry and automated microtiter-based assay. Food Chem. 2008, 111, 707–718.
[23]
Adonizio, A.L.; Downum, K.; Bennett, B.C.; Mathee, K. Anti-quorum sensing activity of medicinal plants in southern Florida. J. Ethnopharmacol. 2006, 105, 427–435.
[24]
Sio, C.F.; Otten, L.G.; Cool, R.H.; Diggle, S.P.; Braun, P.G.; Bos, R.; Daykin, M.; Ca, M.; Williams, P.; Quax, W.J. Quorum quenching by an N-acyl-homoserine lactone acylase from Pseudomonas aeruginosa PAO1. J. Bacteriol. 2006, 74, 1673–1682.
Kloucek, P.; Polesny, Z.; Svobodova, B.; Vlkova, E.; Kokoska, L. Antibacterial screening of some Peruvian medicinal plants used in Callería District. J. Ethnopharmacol. 2005, 99, 309–312.
[27]
Glick, J.; Garber, N. The intracellular localization of Pseudomonas aeruginosa lectins. J. Gen. Microbiol. 1983, 129, 3085–3090.
[28]
Bajolet-Laudinat, O.; Bentzmann, S.G.; Tournier, J.M.; Madoulet, C.; Plotkowski, M.C.; Chippaux, C.; Puchelle, E. Cytotoxicity of Pseudomonas aeruginosa internal lectin PA-I to respiratory epithelial cells in primary culture. Infec. Immun. 1994, 62, 4481–4487.
[29]
Diggle, S.P.; Stacey, R.E.; Dodd, C.; Cámara, M.; Williams, P.; Winzer, K. The galactophilic lectin, LecA, contributes to biofilm development in Pseudomonas aeruginosa. Environ. Microbiol. 2006, 8, 1095–1104.
[30]
Daniels, R.; Vanderleyden, J.; Michiels, J. Quorum sensing and swarming migration in bacteria. FEMS Microbiol. Rev. 2004, 28, 261–289.
[31]
Kearns, D.B. A field guide to bacterial swarming motility. Nat. Rev. Microbiol. 2011, 8, 634–644.
[32]
K?hler, T.; Curty, L.K.; Barja, F.; van Delden, C.; Pechère, J.C. Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J. Bacteriol. 2000, 182, 5990–5996.
[33]
Harshey, R.M. Bees aren't the only ones: Swarming in Gram-negative bacteria. Mol. Microbiol. 1994, 13, 389–394.
[34]
McKnight, S.L.; Iglewski, B.H.; Pesci, E.C. The Pseudomonas quinolone signal regulates rhl quorum sensing in Pseudomonas aeruginosa. J. Bacteriol. 2000, 182, 2702–2708.
[35]
Eberl, L.; Christiansen, G.; Molin, S.; Givskov, M. Differentiation of Serratia liquefaciens into swarm cells is controlled by the expression of the flhD master operon. J. Bacteriol. 1996, 178, 554–559.
[36]
Harshey, R.M.; Matsuyama, T. Dimorphic transition in Escherichia coli and Salmonella typhimurium: Surface-induced differentiation into hyperflagellate swarmer cells. Proc. Natl. Acad. Sci. USA 1994, 91, 8631–8635.
[37]
Lau, G.W.; Ran, H.; Kong, F.; Hassett, D.J.; Mavrodi, D. Pseudomonas aeruginosa pyocyanin is critical for lung infection in mice. Infec. Immun. 2004, 72, 4275–4278.
[38]
Usher, L.R.; Lawson, R.A.; Geary, I.; Taylor, C.J.; Bingle, C.D.; Taylor, G.W.; Whyte, M.K.B. Induction of neutrophil apoptosis by the Pseudomonas aeruginosa exotoxin pyocyanin: A potential mechanism of persistent infection. J. Immunol. 2002, 168, 1861–1868.
[39]
Cao, H.; Krishnan, G.; Goumnerov, B.; Tsongalis, J.; Tompkins, R.; Rahme, L.G. A quorum sensing-associated virulence gene of Pseudomonas aeruginosa encodes a LysR-like transcription regulator with a unique self-regulatory mechanism. Proc. Natl. Acad. Sci. USA 2001, 98, 14613–14618.