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Research article

Molecular and Cellular Microbiology (MCM)  |  Pathogenesis and Host-Microbial interactions

Emodin Successfully Inhibited Invasion of Brucella abortus Via Modulting Adherence, Microtubule Dynamics and ERK Signaling Pathway in RAW 264.7 Cells

Tran Xuan Ngoc Huy , Alisha Wehdnesday Bernardo Reyes , Huynh Tan Hop , Lauren Togonon Arayan , Vu Hai Son , Wongi Min , Hu Jang Lee and Suk Kim *

Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea

Received: April 24, 2018; Accepted: September 1, 2018

J. Microbiol. Biotechnol. 2018; 28(10): 1723-1729

Published October 28, 2018 https://doi.org/10.4014/jmb.1804.04040

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

The aim of this work is to investigate the protective efficacy of emodin, an active, naturallyoccurring anthraquinone derivative of several traditional Chinese herbs, against Brucella abortus infection in macrophages. Brucella were incubated with different concentrations of emodin and showed that bacterial survival rates were markedly reduced in a dose-dependent manner at increasing incubation time points. Through bacterial infection assay, the highest non-cytotoxic concentration of emodin demonstrated attenuated invasion of Brucella into macrophages, however it did not inhibit the growth of these pathogens within the host cells. On the other hand, emodin effectively decreased the number of bacteria that adhered to host cells, which indicated its potential as an anti-adhesin agent. Furthermore, using immunoblotting and FACS assay for detecting MAPK signaling proteins and F-actin polymerization, respectively, the results showed that the emodin-incubated cells displayed modest reduction in the phosphorylation levels of ERK1/2 and inhibition of F-actin polymerization as compared to control cells. These findings indicate the potential use of emodin as a naturally-occurring alternative method for the prevention of animal brucellosis although this requires confirmation of safe clinical doses.

Keywords

Brucella abortus, emodin, antibacterial, invasion

References

  1. Campos MA, Rosinha GMS, Almeida IC, Salgueiro XS, Jarvis BW, Splitter GA, et al. 2004. Role of Toll-like receptor 4 in induction of cell-mediated immunity and resistance to Brucella abortus infection in mice. Infect. Immun. 72: 176-186.
    Pubmed PMC CrossRef
  2. Czyz DM, Jain-Gupta N, Shuman HA, Crosson S. 2016. A dual-targeting approach to inhibit Brucella abortus replication in human cells. Sci. Rep. 6: 35835.
    Pubmed PMC CrossRef
  3. Reyes AWB, Arayan LT, Simborio HL, Hop HT, Min W, Lee HJ, et al. 2016. Dextran sulfate sodium upregulates MAPK signaling for the uptake and subsequent intracellular survival of Brucella abortus in murine macrophages. Microb. Pathog. 91: 68-73.
    Pubmed CrossRef
  4. Wei G, Wu Y, Gao Q, Zhou C, Wang K, Shen C, et al. 2017. Effect of emodin on preventing postoperative intra-abdominal adhesion formation. Oxid. Med. Cell Longev. 2017: 1-12.
    Pubmed PMC CrossRef
  5. Lee JJ, Kim DH, Kim DG, Lee HJ, Min W, Rhee MH, et al. 2012. Phellinus baumii extract influences pathogenesis of Brucella abortus in phagocyte by disrupting the phagocytic and intracellular trafficking pathway. J. Appl. Microbiol. 114:329-338.
    Pubmed CrossRef
  6. Reyes AW, Kim DG, Simborio HL, Hop HT, Arayan LT, Min W, et al. 2016. Methyl gallate limits infection in mice challenged with Brucella abortus while enhancing the inflammatory response. J. Appl. Microbiol. 120: 552-559.
    Pubmed CrossRef
  7. Jimenez de Bagues MP, Gross A, Terraza A, Dornand J. 2005. Regulation of the mitogen-activated protein kinases by Brucella spp. expressing a smooth and rough phenotype:relationship to pathogen invasiveness. Infect. Immun. 73:3178-3183.
    Pubmed PMC CrossRef
  8. Ahmed W, Zheng K, Liu ZF. 2016. Establishment of chronic infection: Brucella’s stealth strategy. Front. Cell Infect. Microbiol. 6: 1-12.
    Pubmed PMC CrossRef
  9. Lee JJ, Kim DH, Kim DG, Lee HJ, Min W, Rhee MH, et al. 2013. Toll-like receptor 4-linked Janus kinase 2 signaling contributes to internalization of Brucella abortus by macrophages. Infect. Immun. 81: 2448-2458.
    Pubmed PMC CrossRef
  10. Wen KW, Bejo SK. 2010. Screening of Chinese medicinal herbs for the inhibition of Brucella melitensis. 5th Proceedings of the Seminar in Veterinary Sciences.
  11. de Figueiredo P, Ficht TA, Rice-Ficht A, Rossetti CA, Adams LG. 2015. Pathogenesis and immunobiology of brucellosis: review of Brucella-host interactions. Am. J. Pathol. 185: 1505-1517.
    Pubmed PMC CrossRef
  12. Lee JJ, Bae JH, Kim DH, Lim JJ, Kim DG, Lee HJ, et al. 2011. Intracellular replication inhibitory effects of Galla Rhois ethanol extract for Brucella abortus infection. J. Ethnopharmacol. 138:602-609.
    Pubmed CrossRef
  13. Stones DH, Krachler AM. 2016. Against the tide: the role of bacterial adhesion in host colonization. Biochem. Soc. Trans. 44: 1571-1580.
    Pubmed PMC CrossRef
  14. Ofek I, Hasty DL, Sharon N. 2003. Anti-adhesion therapy of bacterial diseases: prospects and problems. FEMS Immunol. Med. Microbiol. 38: 181-191.
    CrossRef
  15. Rabin N, Zheng Y, Opoku-Temeng C, Du Y, Bonsu E, Sintim HO. 2015. Agents that inhibit bacterial biofilm formation. Future Med. Chem. 7: 647-671.
    Pubmed CrossRef
  16. Meng G, Liu Y, Lou C, Yang H. 2010. Emodin suppresses lipopolysaccharide-induced pro-inflammatory responses and NF-κB activation by disrupting lipid rafts in CD14-negative endothelial cells. Br. J. Pharmacol. 161: 1628-1644.
    Pubmed PMC CrossRef
  17. Wang HH, Chung JG. 1997. Emodin-induced inhibition of growth and DNA damage in the Helicobacter pylori. Curr. Microbiol. 35: 262-266.
    Pubmed CrossRef
  18. Chukwujekwu JC, Coombes PH, Mulholland DA, van Staden J. 2006. Emodin, an antibacterial anthraquinone from the roots of Cassia occidentalis. S Afr. J. Bot. 72: 295-297.
    CrossRef
  19. Liu M, Peng W, Qin R, Yan Z, Cen Y, Zheng X, et al. 2015. The direct anti-MRSA effect of emodin via damaging cell membrane. Appl. Microbiol. Biotechnol. 99: 7699-7709.
    Pubmed CrossRef
  20. Li L, Song X, Yin Z, Jia R, Li Z, Zhou X, et al. 2016. The antibacterial activity and action mechanism of emodin from Polygonum cuspidatum against Haemophilus parasuis in vitro. Microbiol. Res. 186-187: 139-145.
    Pubmed CrossRef
  21. Pei J, Turse JE, Ficht TA. 2008. Evidence of Brucella abortus OPS dictating uptake and restricting NF-kB activation in murine macrophages. Microbes Infect. 10: 582-590.
    Pubmed PMC CrossRef
  22. Huang Q, Shen HM, Ong CN. 2005. Emodin inhibits tumor cell migration through suppression of the phosphatidylinositol 3-kinase-Cdc42/Rac1 pathway. Cell Mol. Life Sci. 62: 1167-1175.
    Pubmed CrossRef
  23. Shan L, He P, Sheen J. 2007. Intercepting host MAPK signaling cascades by bacterial type III effectors. Cell Host Microbe 1: 167-174.
    Pubmed CrossRef
  24. Mijatovic S, Maksimovic-Ivanic D, Radovic J, Miljkovic D, Harhaji L, Vuckovic O, et al. 2005. Anti-glioma action of aloe emodin: the role of ERK inhibition. Cell Mol. Life Sci. 62: 589-598.
    Pubmed CrossRef
  25. Cui Y, Lu P, Song G, Liu Q, Zhu D, Liu X. 2016. Involvement of PI3K/Akt, ERK and p38 signaling pathways in emodin-mediated extrinsic and intrinsic human hepatoblastoma cell apoptosis. Food Chem. Toxicol. 92: 26-37.
    Pubmed CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2018; 28(10): 1723-1729

Published online October 28, 2018 https://doi.org/10.4014/jmb.1804.04040

Copyright © The Korean Society for Microbiology and Biotechnology.

Emodin Successfully Inhibited Invasion of Brucella abortus Via Modulting Adherence, Microtubule Dynamics and ERK Signaling Pathway in RAW 264.7 Cells

Tran Xuan Ngoc Huy , Alisha Wehdnesday Bernardo Reyes , Huynh Tan Hop , Lauren Togonon Arayan , Vu Hai Son , Wongi Min , Hu Jang Lee and Suk Kim *

Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea

Received: April 24, 2018; Accepted: September 1, 2018

Abstract

The aim of this work is to investigate the protective efficacy of emodin, an active, naturallyoccurring
anthraquinone derivative of several traditional Chinese herbs, against Brucella
abortus infection in macrophages. Brucella were incubated with different concentrations of
emodin and showed that bacterial survival rates were markedly reduced in a dose-dependent
manner at increasing incubation time points. Through bacterial infection assay, the highest
non-cytotoxic concentration of emodin demonstrated attenuated invasion of Brucella into
macrophages, however it did not inhibit the growth of these pathogens within the host cells.
On the other hand, emodin effectively decreased the number of bacteria that adhered to host
cells, which indicated its potential as an anti-adhesin agent. Furthermore, using
immunoblotting and FACS assay for detecting MAPK signaling proteins and F-actin
polymerization, respectively, the results showed that the emodin-incubated cells displayed
modest reduction in the phosphorylation levels of ERK1/2 and inhibition of F-actin
polymerization as compared to control cells. These findings indicate the potential use of
emodin as a naturally-occurring alternative method for the prevention of animal brucellosis
although this requires confirmation of safe clinical doses.

Keywords: Brucella abortus, emodin, antibacterial, invasion

References

  1. Campos MA, Rosinha GMS, Almeida IC, Salgueiro XS, Jarvis BW, Splitter GA, et al. 2004. Role of Toll-like receptor 4 in induction of cell-mediated immunity and resistance to Brucella abortus infection in mice. Infect. Immun. 72: 176-186.
    Pubmed KoreaMed CrossRef
  2. Czyz DM, Jain-Gupta N, Shuman HA, Crosson S. 2016. A dual-targeting approach to inhibit Brucella abortus replication in human cells. Sci. Rep. 6: 35835.
    Pubmed KoreaMed CrossRef
  3. Reyes AWB, Arayan LT, Simborio HL, Hop HT, Min W, Lee HJ, et al. 2016. Dextran sulfate sodium upregulates MAPK signaling for the uptake and subsequent intracellular survival of Brucella abortus in murine macrophages. Microb. Pathog. 91: 68-73.
    Pubmed CrossRef
  4. Wei G, Wu Y, Gao Q, Zhou C, Wang K, Shen C, et al. 2017. Effect of emodin on preventing postoperative intra-abdominal adhesion formation. Oxid. Med. Cell Longev. 2017: 1-12.
    Pubmed KoreaMed CrossRef
  5. Lee JJ, Kim DH, Kim DG, Lee HJ, Min W, Rhee MH, et al. 2012. Phellinus baumii extract influences pathogenesis of Brucella abortus in phagocyte by disrupting the phagocytic and intracellular trafficking pathway. J. Appl. Microbiol. 114:329-338.
    Pubmed CrossRef
  6. Reyes AW, Kim DG, Simborio HL, Hop HT, Arayan LT, Min W, et al. 2016. Methyl gallate limits infection in mice challenged with Brucella abortus while enhancing the inflammatory response. J. Appl. Microbiol. 120: 552-559.
    Pubmed CrossRef
  7. Jimenez de Bagues MP, Gross A, Terraza A, Dornand J. 2005. Regulation of the mitogen-activated protein kinases by Brucella spp. expressing a smooth and rough phenotype:relationship to pathogen invasiveness. Infect. Immun. 73:3178-3183.
    Pubmed KoreaMed CrossRef
  8. Ahmed W, Zheng K, Liu ZF. 2016. Establishment of chronic infection: Brucella’s stealth strategy. Front. Cell Infect. Microbiol. 6: 1-12.
    Pubmed KoreaMed CrossRef
  9. Lee JJ, Kim DH, Kim DG, Lee HJ, Min W, Rhee MH, et al. 2013. Toll-like receptor 4-linked Janus kinase 2 signaling contributes to internalization of Brucella abortus by macrophages. Infect. Immun. 81: 2448-2458.
    Pubmed KoreaMed CrossRef
  10. Wen KW, Bejo SK. 2010. Screening of Chinese medicinal herbs for the inhibition of Brucella melitensis. 5th Proceedings of the Seminar in Veterinary Sciences.
  11. de Figueiredo P, Ficht TA, Rice-Ficht A, Rossetti CA, Adams LG. 2015. Pathogenesis and immunobiology of brucellosis: review of Brucella-host interactions. Am. J. Pathol. 185: 1505-1517.
    Pubmed KoreaMed CrossRef
  12. Lee JJ, Bae JH, Kim DH, Lim JJ, Kim DG, Lee HJ, et al. 2011. Intracellular replication inhibitory effects of Galla Rhois ethanol extract for Brucella abortus infection. J. Ethnopharmacol. 138:602-609.
    Pubmed CrossRef
  13. Stones DH, Krachler AM. 2016. Against the tide: the role of bacterial adhesion in host colonization. Biochem. Soc. Trans. 44: 1571-1580.
    Pubmed KoreaMed CrossRef
  14. Ofek I, Hasty DL, Sharon N. 2003. Anti-adhesion therapy of bacterial diseases: prospects and problems. FEMS Immunol. Med. Microbiol. 38: 181-191.
    CrossRef
  15. Rabin N, Zheng Y, Opoku-Temeng C, Du Y, Bonsu E, Sintim HO. 2015. Agents that inhibit bacterial biofilm formation. Future Med. Chem. 7: 647-671.
    Pubmed CrossRef
  16. Meng G, Liu Y, Lou C, Yang H. 2010. Emodin suppresses lipopolysaccharide-induced pro-inflammatory responses and NF-κB activation by disrupting lipid rafts in CD14-negative endothelial cells. Br. J. Pharmacol. 161: 1628-1644.
    Pubmed KoreaMed CrossRef
  17. Wang HH, Chung JG. 1997. Emodin-induced inhibition of growth and DNA damage in the Helicobacter pylori. Curr. Microbiol. 35: 262-266.
    Pubmed CrossRef
  18. Chukwujekwu JC, Coombes PH, Mulholland DA, van Staden J. 2006. Emodin, an antibacterial anthraquinone from the roots of Cassia occidentalis. S Afr. J. Bot. 72: 295-297.
    CrossRef
  19. Liu M, Peng W, Qin R, Yan Z, Cen Y, Zheng X, et al. 2015. The direct anti-MRSA effect of emodin via damaging cell membrane. Appl. Microbiol. Biotechnol. 99: 7699-7709.
    Pubmed CrossRef
  20. Li L, Song X, Yin Z, Jia R, Li Z, Zhou X, et al. 2016. The antibacterial activity and action mechanism of emodin from Polygonum cuspidatum against Haemophilus parasuis in vitro. Microbiol. Res. 186-187: 139-145.
    Pubmed CrossRef
  21. Pei J, Turse JE, Ficht TA. 2008. Evidence of Brucella abortus OPS dictating uptake and restricting NF-kB activation in murine macrophages. Microbes Infect. 10: 582-590.
    Pubmed KoreaMed CrossRef
  22. Huang Q, Shen HM, Ong CN. 2005. Emodin inhibits tumor cell migration through suppression of the phosphatidylinositol 3-kinase-Cdc42/Rac1 pathway. Cell Mol. Life Sci. 62: 1167-1175.
    Pubmed CrossRef
  23. Shan L, He P, Sheen J. 2007. Intercepting host MAPK signaling cascades by bacterial type III effectors. Cell Host Microbe 1: 167-174.
    Pubmed CrossRef
  24. Mijatovic S, Maksimovic-Ivanic D, Radovic J, Miljkovic D, Harhaji L, Vuckovic O, et al. 2005. Anti-glioma action of aloe emodin: the role of ERK inhibition. Cell Mol. Life Sci. 62: 589-598.
    Pubmed CrossRef
  25. Cui Y, Lu P, Song G, Liu Q, Zhu D, Liu X. 2016. Involvement of PI3K/Akt, ERK and p38 signaling pathways in emodin-mediated extrinsic and intrinsic human hepatoblastoma cell apoptosis. Food Chem. Toxicol. 92: 26-37.
    Pubmed CrossRef