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In Vitro Antimicrobial Activity and Downregulation of Virulence Gene Expression on Helicobacter pylori by Reuterin

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Abstract

Helicobacter pylori is an infectious agent commonly associated with gastrointestinal diseases. The use of probiotics to treat this infection has been documented, however, their potential antimicrobial metabolites have not yet been investigated. In the present study, the effect of reuterin produced by Lactobacillus reuteri on H. pylori growth and virulence gene expression was evaluated. It was observed that reuterin caused significant (P < 0.05) H. pylori growth inhibition at concentrations from 0.08 to 20.48 mM, with minimal inhibitory concentrations (MICs) of 20.48 mM for H. pylori ATCC700824 and 10.24 mM for H. pylori ATCC43504. In a reuterin bacterial killing assay, it was observed that half of the MIC value for H. pylori (ATCC700824) significantly (P < 0.01) reduced colony numbers from 5.65 ± 0.35 to 3.78 ± 0.35 Log10 CFU/mL after 12 h of treatment and then increased them to 5.25 ± 0.23 Log10 CFU/mL at 24 h; at its MIC value (20.48 mM), reuterin abrogated (P < 0.01) H. pylori (ATCC700824) growth after 20 h of culture. In addition, reuterin significantly (P < 0.01) reduced H. pylori (ATCC 43504) colony numbers from 5.65 ± 0.35 to 4.1 ± 0.12 Log10 CFU/mL from 12 to 24 h of treatment and abrogated its growth at its MIC value (10.24 mM), after 20 h of treatment. Reuterin did not alter normal human gastric Hs738.St/Int cell viability at the concentrations tested for H. pylori strains. Furthermore, 10 μM reuterin was shown to significantly (P < 0.01) reduce mRNA relative expression levels of H. pylori virulence genes vacA and flaA at 3 h post-treatment, whose effect was higher at 6 h post-treatment, as measured by RT-qPCR. The observed direct antimicrobial effect and the downregulation of expression of virulence genes on H. pylori by reuterin may contribute to the understanding of the mechanisms of action of probiotics against H. pylori.

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Acknowledgements

We thank Laboratorio de Inmunología y Virología of Facultad de Ciencias Biológicas at Universidad Autónoma de Nuevo León for supporting the development of this study.

Funding

This study was partly supported by grants from Programa de Apoyo a la Investigación Científica y Tecnológica from Universidad Autónoma de Nuevo León (PAICYT-UANL) to RGF and by Consejo Nacional de Ciencia y Tecnología (CONACYT-México) to VUB.

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Authors and Affiliations

  1. Universidad Autónoma de Nuevo León, Facultad de Ciencias Biológicas, Departamento de Microbiología e Inmunología, Laboratorio de Inmunología y Virología, Pedro de Alba y Manuel L. Barragán S/N. Cd. Universitaria, CP. 66450, San Nicolás de los Garza, N.L., México

    Víctor Hugo Urrutia-Baca, Patricia Tamez-Guerra, Ricardo Gomez-Flores & Cynthia Sofía Urbina-Ríos

  2. Universidad Autónoma de Nuevo León, Facultad de Odontología y Unidad de Odontología Integral y Especialidades del CIDICS, Av. Dr. Aguirre Pequeño y Silao S/N, Colonia Mitras Centro., CP. 64460, Monterrey, N.L., México

    Erandi Escamilla-García & Myriam Angélica de la Garza-Ramos

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  1. Víctor Hugo Urrutia-Baca
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  2. Erandi Escamilla-García
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Correspondence to Ricardo Gomez-Flores.

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Urrutia-Baca, V.H., Escamilla-García, E., de la Garza-Ramos, M.A. et al. In Vitro Antimicrobial Activity and Downregulation of Virulence Gene Expression on Helicobacter pylori by Reuterin. Probiotics & Antimicro. Prot. 10, 168–175 (2018). https://doi.org/10.1007/s12602-017-9342-2

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