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Down-regulatory effects of green coffee extract on las I and las R virulence-associated genes in Pseudomonas aeruginosa

Abstract

Background

Antibiotic resistant strains of Pseudomonas aeruginosa are the cause of Gram negative nosocomial infections especially among the immunosuppressed patients. The bacteria contains las I and las R genes that play very important roles in the pathogenesis and mechanisms of aggression. These genes can be influenced by the quorum sensing (QS) system and such mechanism is becoming clinically important worldwide. This study aimed to investigate the preventive effects of green coffee extract (GCE) on the expression of pathogenesis-related genes, las I and las R in P. aeruginosa.

Methods

A total of fifty four P. aeruginosa strains were isolated out of 100 clinical samples collected from the infectious wards in different hospitals (Tehran province) using conventional microscopic and biochemical methods. Susceptibility of the isolates to different antibiotics, GCE and chlorogenic acid were elucidated. Multiplex polymerase chain reaction (PCR) and real-time PCR were performed to detect and quantify the expression levels of las I and las R genes. The presence of chlorogenic acid in GCE was confirmed by HPLC.

Results

Antibiotic susceptibility tests revealed multidrug resistance among the clinical isolates of those 40 strains were resistant to ciprofloxacin (74.07%), 43 to ceftazidime (79.26%), 29 to amikacin (53.7%), 42 to ampicillin (77.77%), 17 to colistin (31.48%), 40 to gentamicin (74.77%), and 50 to piperacillin (92.59%). PCR outcomes exhibited that the frequency of las I and las R genes were 100% in resistant and sensitive strains isolated from clinical and standard strains of P. aeruginosa (ATCC 15449). Real-time PCR analyses revealed that GCE significantly prevented the expression of las I and las R genes in P. aeruginosa. GCE at concentration level as low as 2.5 mg/mL could prevent the expression of lasI and lasR genes in P. aeruginosa clinical isolates.

Conclusion

The presence and expression levels of las I and las R genes in P. aeruginosa isolates were investigated when the bacteria was exposed to GCE. Our results tend to suggest that genes involved in pathogenesis of:Pseudomonas aeruginosa are down regulated by quorum sensing effect of chlorogenic acid and therefore GCE could be useful as an adjuvant in combating multidrug resistance strains of Pseudomonas aeruginosa.

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References

  1. 1.

    Brooks G, Carroll KC, Butel J, Morse S. Jawetz Melnick & Adelbergs Medical Microbiology 26/E: McGraw Hill Professional; 2012.

  2. 2.

    Karatuna O, Yagci A. Analysis of quorum sensing-dependent virulence factor production and its relationship with antimicrobial susceptibility in Pseudomonas aeruginosa respiratory isolates. Clin Microbiol Infect. 2010;16(12):1770–5.

    Article  CAS  PubMed  Google Scholar 

  3. 3.

    Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP. The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science. 1998;280(5361):295–8.

    Article  CAS  PubMed  Google Scholar 

  4. 4.

    Annapoorani A, Umamageswaran V, Parameswari R, Pandian SK, Ravi AV. Computational discovery of putative quorum sensing inhibitors against LasR and RhlR receptor proteins of Pseudomonas aeruginosa. J Comput Aided Mol Des. 2012;26(9):1067–77.

    Article  CAS  PubMed  Google Scholar 

  5. 5.

    Zhu H, Conibear TC, Bandara R, Aliwarga Y, Stapleton F, Willcox MD. Type III secretion system–associated toxins, proteases, serotypes, and antibiotic resistance of Pseudomonas aeruginosa isolates associated with keratitis. Curr Eye Res. 2006;31(4):297–306.

    Article  CAS  PubMed  Google Scholar 

  6. 6.

    Waters CM, Bassler BL. Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol. 2005;21:319–46.

    Article  CAS  Google Scholar 

  7. 7.

    Wagner VE, Frelinger JG, Barth RK, Iglewski BH. Quorum sensing: dynamic response of Pseudomonas aeruginosa to external signals. Trends Microbiol. 2006;14(2):55–8.

    Article  CAS  PubMed  Google Scholar 

  8. 8.

    Reading NC, Sperandio V. Quorum sensing: the many languages of bacteria. FEMS Microbiol Lett. 2005;254(1):1–11.

    Article  CAS  Google Scholar 

  9. 9.

    Raina S, Vizio DD, Odell M, Clements M, Vanhulle S, Keshavarz T. Microbial quorum sensing: a tool or a target for antimicrobial therapy? Biotechnol Appl Biochem. 2009;54(2):65–84.

    Article  CAS  PubMed  Google Scholar 

  10. 10.

    Adonizio A, Kong K-F, Mathee K. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by South Florida plant extracts. Antimicrob Agents Chemother. 2008;52(1):198–203.

    Article  CAS  PubMed  Google Scholar 

  11. 11.

    Fratianni F, Pepe R, Nazzaro F. Polyphenol composition, antioxidant, antimicrobial and quorum quenching activity of the “Carciofo di Montoro”(Cynara cardunculus var. scolymus) global artichoke of the Campania region, southern Italy. Food Nutr Sci. 2014;5(21):2053–62.

    CAS  Google Scholar 

  12. 12.

    Kalia VC. Quorum sensing inhibitors: an overview. Biotechnol Adv. 2013;31(2):224–45.

    Article  CAS  PubMed  Google Scholar 

  13. 13.

    Santana-Gálvez J, Cisneros-Zevallos L, Jacobo-Velázquez DA. Chlorogenic acid: recent advances on its dual role as a food additive and a nutraceutical against metabolic syndrome. Molecules. 2017;22(3):358.

    Article  CAS  PubMed Central  Google Scholar 

  14. 14.

    Kozuma K, Tsuchiya S, Kohori J, Hase T, Tokimitsu I. Antihypertensive effect of green coffee bean extract on mildly hypertensive subjects. Hypertens Res. 2005;28(9):711–8.

    Article  CAS  PubMed  Google Scholar 

  15. 15.

    Ochiai R, Jokura H, Suzuki A, Tokimitsu I, Ohishi M, Komai N, et al. Green coffee bean extract improves human vasoreactivity. Hypertens Res. 2004;27(10):731–7.

    Article  CAS  PubMed  Google Scholar 

  16. 16.

    Shimoda H, Seki E, Aitani M. Inhibitory effect of green coffee bean extract on fat accumulation and body weight gain in mice. BMC Complement Altern Med. 2006;6(1):9.

    Article  PubMed  PubMed Central  Google Scholar 

  17. 17.

    Suzuki A, Kagawa D, Ochiai R, Tokimitsu I, Saito I. Green coffee bean extract and its metabolites have a hypotensive effect in spontaneously hypertensive rats. Hypertens Res. 2002;25(1):99–107.

    Article  CAS  PubMed  Google Scholar 

  18. 18.

    Blum J, Lemaire B, Lafay S. Effect of a green decaffeinated coffee extract on glycaemia. NutraFoods 2007;6:13–7.

    CAS  Google Scholar 

  19. 19.

    Farah A, Donangelo CM. Phenolic compounds in coffee. Braz J Plant Physiol. 2006;18(1):23–36.

    Article  CAS  Google Scholar 

  20. 20.

    Suárez-Quiroz ML, Taillefer W, López Méndez EM, González-Ríos O, Villeneuve P, Figueroa-Espinoza M-C. Antibacterial activity and antifungal and anti-Mycotoxigenic activities against aspergillus flavus and A. ochraceus of green coffee Chlorogenic acids and dodecyl Chlorogenates. J Food Saf. 2013;33(3):360–8.

    Article  CAS  Google Scholar 

  21. 21.

    Garrity GM. Bergey's manual of systematic bacteriology: volume one: the archaea and the deeply branching and phototrophic Bacteria: Springer Science & Business Media; 2012.

  22. 22.

    Almeida AAP, Farah A, Silva DA, Nunan EA, MBA G. Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria. J Agric Food Chem. 2006;54(23):8738–43.

    Article  CAS  PubMed  Google Scholar 

  23. 23.

    Ayelign A, Sabally K. Determination of chlorogenic acids (CGA) in coffee beans using HPLC. Am J Res Commun. 2013;1(2):78–91.

    Google Scholar 

  24. 24.

    Kadhim D, Ali MR. Prevalence study of quorum sensing groups among clinical isolates of Pseudomonas aeruginosa. Int J Curr Microbiol App Sci. 2014;3(11):204–15.

    CAS  Google Scholar 

  25. 25.

    Thuptimdang P, Limpiyakorn T, McEvoy J, Prüß BM, Khan E. Effect of silver nanoparticles on Pseudomonas putida biofilms at different stages of maturity. J Hazard Mater. 2015;290:127–33.

    Article  CAS  PubMed  Google Scholar 

  26. 26.

    Hoque MM, Ahmad M, Khisa S, Uddin MN, Jesmine R. Antibiotic resistance pattern in Pseudomonas aeruginosa isolated from different clinical specimens. JAFMC. 2016;11(1):45–9.

    Google Scholar 

  27. 27.

    Singh Arora D, Jeet Kaur G, Kaur H. Antibacterial activity of tea and coffee: their extracts and preparations. Int J Food Prop. 2009;12(2):286–94.

    Article  Google Scholar 

  28. 28.

    Sepahi E, Tarighi S, Ahmadi FS, Bagheri A. Inhibition of quorum sensing in Pseudomonas aeruginosa by two herbal essential oils from Apiaceae family. J Microbiol. 2015;53(2):176–80.

    Article  CAS  PubMed  Google Scholar 

  29. 29.

    Senturk S, Ulusoy S, Bosgelmez-Tinaz G, Yagci A. Quorum sensing and virulence of Pseudomonas aeruginosa during urinary tract infections. The Journal of Infection in Developing Countries. 2012;6(06):501–7.

    CAS  Article  Google Scholar 

  30. 30.

    Farah A, Monteiro M, Donangelo CM, Lafay S. Chlorogenic acids from green coffee extract are highly bioavailable in humans. J Nutr. 2008;138(12):2309–15.

    Article  CAS  PubMed  Google Scholar 

  31. 31.

    Ranheim T, Halvorsen B. Coffee consumption and human health–beneficial or detrimental?–mechanisms for effects of coffee consumption on different risk factors for cardiovascular disease and type 2 diabetes mellitus. Mol Nutr Food Res. 2005;49(3):274–84.

    Article  CAS  PubMed  Google Scholar 

  32. 32.

    Lindsay J, Laurin D, Verreault R, Hébert R, Helliwell B, Hill GB, et al. Risk factors for Alzheimer’s disease: a prospective analysis from the Canadian study of health and aging. Am J Epidemiol. 2002;156(5):445–53.

    Article  PubMed  Google Scholar 

  33. 33.

    Craig AP, Fields C, Liang N, Kitts D, Erickson A. Performance review of a fast HPLC-UV method for the quantification of chlorogenic acids in green coffee bean extracts. Talanta. 2016;154:481–5.

    Article  CAS  PubMed  Google Scholar 

  34. 34.

    Karunanidhi A, Thomas R, Van Belkum A, Neela V. In vitro antibacterial and antibiofilm activities of chlorogenic acid against clinical isolates of Stenotrophomonas maltophilia including the trimethoprim/sulfamethoxazole resistant strain. Biomed Res Int. 2012;2013.

  35. 35.

    Kabir F, Katayama S, Tanji N, Nakamura S. Antimicrobial effects of chlorogenic acid and related compounds. J Korean Soc Appl Biol Chem. 2014;57(3):359–65.

    Article  CAS  Google Scholar 

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Funding

This research was supported by Deputy of Research, Tehran University of Medical Sciences, (grant No. 31002).

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Authors

Contributions

This study was PhD thesis of HJ who did all the practical experiments. HJ contributed in the conception, design of the study, data collation and manuscript preparation. NS contributed in the conception and the design of the study. JN contributed in the design of the study. MD supervised the gene analysis experiments. MRF contributed in the conception, the design of the study and gave revisions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mohammad Reza Fazeli.

Ethics declarations

This study was approved by the Ethics Committee of the Tehran University of Medical Sciences, Tehran, Iran (ethics committee reference number: IR. TUMS. REC. 1394.2108).

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The authors declare that they have no competing interest.

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Jamalifar, H., Samadi, N., Nowroozi, J. et al. Down-regulatory effects of green coffee extract on las I and las R virulence-associated genes in Pseudomonas aeruginosa. DARU J Pharm Sci 27, 35–42 (2019). https://doi.org/10.1007/s40199-018-0234-0

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Keywords

  • Pseudomonas aeruginosa
  • Chlorogenic acid
  • Green coffee extract
  • Quorum sensing