DOI QR코드

DOI QR Code

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

  • Huy, Tran Xuan Ngoc (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Reyes, Alisha Wehdnesday Bernardo (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Hop, Huynh Tan (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Arayan, Lauren Togonon (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Son, Vu Hai (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Min, Wongi (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Lee, Hu Jang (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Suk (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
  • Received : 2018.04.24
  • Accepted : 2018.09.01
  • Published : 2018.10.28

Abstract

The aim of this work is to investigate the protective efficacy of emodin, an active, naturally-occurring 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

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. https://doi.org/10.1128/IAI.72.1.176-186.2004
  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. https://doi.org/10.1038/srep35835
  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. https://doi.org/10.1016/j.micpath.2015.10.024
  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.
  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.
  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. https://doi.org/10.1111/jam.13019
  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. https://doi.org/10.1128/IAI.73.5.3178-3183.2005
  8. Ahmed W, Zheng K, Liu ZF. 2016. Establishment of chronic infection: Brucella's stealth strategy. Front. Cell Infect. Microbiol. 6: 1-12.
  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. https://doi.org/10.1128/IAI.00403-13
  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. https://doi.org/10.1016/j.ajpath.2015.03.003
  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. https://doi.org/10.1016/j.jep.2011.10.007
  13. Stones DH, Krachler AM. 2016. Against the tide: the role of bacterial adhesion in host colonization. Biochem. Soc. Trans. 44: 1571-1580. https://doi.org/10.1042/BST20160186
  14. Ofek I, Hasty DL, Sharon N. 2003. Anti-adhesion therapy of bacterial diseases: prospects and problems. FEMS Immunol. Med. Microbiol. 38: 181-191. https://doi.org/10.1016/S0928-8244(03)00228-1
  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. https://doi.org/10.4155/fmc.15.7
  16. Meng G, Liu Y, Lou C, Yang H. 2010. Emodin suppresses lipopolysaccharide-induced pro-inflammatory responses and $NF-{\kappa}B$ activation by disrupting lipid rafts in CD14-negative endothelial cells. Br. J. Pharmacol. 161: 1628-1644. https://doi.org/10.1111/j.1476-5381.2010.00993.x
  17. Wang HH, Chung JG. 1997. Emodin-induced inhibition of growth and DNA damage in the Helicobacter pylori. Curr. Microbiol. 35: 262-266. https://doi.org/10.1007/s002849900250
  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. https://doi.org/10.1016/j.sajb.2005.08.003
  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. https://doi.org/10.1007/s00253-015-6657-3
  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. https://doi.org/10.1016/j.micres.2016.03.008
  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. https://doi.org/10.1016/j.micinf.2008.01.005
  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. https://doi.org/10.1007/s00018-005-5050-2
  23. Shan L, He P, Sheen J. 2007. Intercepting host MAPK signaling cascades by bacterial type III effectors. Cell Host Microbe 1: 167-174. https://doi.org/10.1016/j.chom.2007.04.008
  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. https://doi.org/10.1007/s00018-005-4425-8
  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. https://doi.org/10.1016/j.fct.2016.03.013

Cited by

  1. Roles of Protein Histidine Phosphatase 1 (PHPT1) in Brown Adipocyte Differentiation vol.30, pp.2, 2018, https://doi.org/10.4014/jmb.1909.09003
  2. Comparative transcriptomic and proteomic analysis of the antibacterial activity of emodin on Aeromonas hydrophila vol.529, pp.None, 2018, https://doi.org/10.1016/j.aquaculture.2020.735589