Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지
DOI QR코드

DOI QR Code

Ursolic Acid Reduces Mycobacterium tuberculosis-Induced Nitric Oxide Release in Human Alveolar A549 cells

  • Zerin, Tamanna (Department of Microbiology, School of Medicine, Soonchunhyang University) ;
  • Lee, Minjung (Department of Microbiology, School of Medicine, Soonchunhyang University) ;
  • Jang, Woong Sik (Regional Innovation Center, Soonchunhyang University) ;
  • Nam, Kung-Woo (Department of Life Science and Biotechnology, College of Natural Science, Soonchunhyang University) ;
  • Song, Ho-yeon (Department of Microbiology, School of Medicine, Soonchunhyang University)
  • Received : 2014.11.26
  • Accepted : 2015.05.14
  • Published : 2015.07.31
Download PDF
⟨ Previous Next ⟩

Abstract

Alveolar epithelial cells have been functionally implicated in Mycobacterium tuberculosis infection. This study investigated the role of ursolic acid (UA)-a triterpenoid carboxylic acid with potent antioxidant, anti-tumor, anti-inflammatory, and anti-tuberculosis properties in mycobacterial infection of alveolar epithelial A549 cells. We observed that M. tuberculosis successfully entered A549 cells. Cytotoxicity was mediated by nitric oxide (NO). A549 toxicity peaked along with NO generation 72 h after infection. The NO generated by mycobacterial infection in A549 cells was insufficient to kill mycobacteria, as made evident by the mycobacteria growth indicator tube time to detect (MGIT TTD) and viable cell count assays. Treatment of mycobacteria-infected cells with UA reduced the expression of inducible nitric oxide synthase, NO generation, and eventually improved cell viability. Moreover, UA was found to quench the translocation of the transcription factor, nuclear factor kappa B (NF-${\kappa}B$), from the cytosol to the nucleus in mycobacteria-infected cells. This study is the first to demonstrate the cytotoxic role of NO in the eradication of mycobacteria and the role of UA in reducing this cytotoxicity in A549 cells.

Keywords

References

  1. Bermudez, L.E., and Young, L.S. (1994). Factors affecting invasion of HT-29 and HEp-2 epithelial cells by organisms of the Mycobacterium avium complex. Infect Immun. 62, 2021-2026.
  2. Bermudez, L.E., and Goodman, J. (1996). Mycobacterium tuberculosis invades and replicates within type II alveolar cells. Infect Immun. 64, 1400-1406.
  3. Chan, J., Xing, Y., Magliozzo, R., and Bloom, B.R. (1992). Killing of virulent Mycobacterium tuberculosis by reactive nitrogen intermediates produced by activated murine macrophages. J. Exp. Med. 175, 1111-1122. https://doi.org/10.1084/jem.175.4.1111
  4. Danelishvili L., McGarvey J., Li Y.J., and Bermudez L.E. (2003). Mycobacterium tuberculosis infection causes different levels of apoptosis and necrosis in human macrophages and alveolar epithelial cells. Cell Microbiol. 5, 649-660. https://doi.org/10.1046/j.1462-5822.2003.00312.x
  5. De las Heras, B., Rodri guez, B., Bosca, L., and Villar, A.M. (2003). Terpenoids: sources, structure elucidation and therapeutic potential in inflammation. Curr. Top Med. Chem. 3, 171-185. https://doi.org/10.2174/1568026033392462
  6. Dhiman, R., Raje, M., and Majumdar, S. (2007). Differential expression of NF-kappaB in mycobacteria infected THP-1 affects apoptosis. Biochim. Biophys. Acta 1770, 649-658. https://doi.org/10.1016/j.bbagen.2006.11.016
  7. Eizirik, D.L., Flodstrom, M., Karlsen, A.E., and Welsh, N. (1996). The harmony of the spheres: Inducible nitric oxide synthase and related genes in pancreatic beta cells. Diabetologia 39, 875-890.
  8. Ellner, J.J. (1997). Review: The immune response in tuberculosis: implication for tuberculosis control. J Infect Dis. 176, 1351-1359. https://doi.org/10.1086/514132
  9. Fenton, M.J., and Vermeulen, M.W. (1996). Immunopathology of tuberculosis: Roles of macrophages and monocytes. Infect Immun. 64, 683-690.
  10. Fulton, S.A., Cross, J.V., Toossi, Z.T., and Boom, W.H. (1998). Regulation of interleukin-12 by interleukin 10, transforming growth factor-b, tumour necrosis factor-a and interferon-c in human monocytes infected with Mycobacterium tuberculosis H37Ra. J. Infect Dis. 178, 1105-1114. https://doi.org/10.1086/515698
  11. Garcia-Perez, B.E., Mondragon-Flores, R., and Luna-Herrera, J. (2003). internalization of Mycobacterium tuberculosis by macropinocytosis in non-phagocytic cells. Microb. Pathog. 35, 49-55. https://doi.org/10.1016/S0882-4010(03)00089-5
  12. Gribar, S.C., Anand, R.J., Sodhi, C.P., and Hackam, D.J. (2008). The role of epithelial Toll-like receptor signaling in the pathogenesis of intestinal inflammation. J. Leukoc. Biol. 83, 493-498. https://doi.org/10.1189/jlb.0607358
  13. Guzik, T.J., Korbut, R., and Adamek-Guzik, T. (2003). Nitric oxide and superoxide in inflammation and immune regulation. J. Physiol Pharmacol. 54, 469-487.
  14. Ignarro, L.J. (2000). Introduction and overview, in Nitric Oxide, Biology and Pathobiology, ed. Ignarro, L.J. (Academic, San Diego), pp 3-19.
  15. Ikeda, Y., Murakami, A., and Ohigashi, H. (2008). Ursolic acid: an anti-and proinflammatory triterpenoid. Mol. Nutr. Food Res. 52, 26-42. https://doi.org/10.1002/mnfr.200700389
  16. Jimenez-Arellanes, A., Luna-Herrera, J., Cornejo-Garrido, J., Lopez-Garcia, S., Castro-Mussot, M.E., Meckes-Fischer, M., Mata-Espinosa, D., Marquina, B., Torres, J., and Hernandez-Pando, R. (2013). Ursolic and oleanolic acids as antimicrobial and immunomodulatory compounds for tuberculosis treatment. BMC Complement. Altern. Med. 13, 258. https://doi.org/10.1186/1472-6882-13-258
  17. Kim, Y.S., Zerin, T., and Song, H.Y. (2013). Antioxidant action of ellagic acid ameliorates paraquat-induced A549 cytotoxicity. Biol. Pharm. Bull. 36, 609-615. https://doi.org/10.1248/bpb.b12-00990
  18. Liu, J. (1995). Pharmacology of oleanolic acid and ursolic acid. J. Ethnopharmacol. 49, 57-68. https://doi.org/10.1016/0378-8741(95)90032-2
  19. Mapother, M.E., and Sanger, J.G. (1984). In vitro interaction of Mycobacterium avium with intestinal epithelial cells. Infect Immun. 45, 67-73.
  20. Murphy, M.P. (1999). Nitric oxide and cell death. Biochim. Biophys. Acta 1411, 401-414. https://doi.org/10.1016/S0005-2728(99)00029-8
  21. Podder, B., Jang, W.S., Nam, K.W., Lee, B.E., and Song, H.Y. (2015). Ursolic acid activates intracellular killing effect of macrophage in Mycobacterium tuberculosis infection. J. Microbiol. Biotechnol. 25, 738-744. https://doi.org/10.4014/jmb.1407.07020
  22. Rich, E.A., Torres, M., Sada, E., Finegan, C.K., Hamilton, B.D., and Toossi, Z. (1997). Mycobacterium tuberculosis stimulated production of nitric oxide by human alveolar macrophages and relationship of nitric oxide production to growth inhibition of MTB. Tubercle Lung Dis. 78, 247-255. https://doi.org/10.1016/S0962-8479(97)90005-8
  23. Roy, S., Sharma, S., Sharma, M., Aggarwal, R., and Bose, M. (2004). Induction of nitric oxide release from the human alveolar epithelial cell line A549: an in vitro correlate of innate immune response to Mycobacterium tuberculosis. Immunology 112, 471-480. https://doi.org/10.1046/j.1365-2567.2004.01905.x
  24. Shepard, C.C. (1955). Phagocytosis by HeLa cells and their susceptibility to infection by human tubercle bacilli. Proc. Soc. Exp. Biol. Med. 90, 392-396. https://doi.org/10.3181/00379727-90-22043
  25. Shishodia, S., Majumdar, S., Banerjee, S., and Aggarwal, B.B. (2003). Ursolic acid inhibits nuclear factor-kappaB activation induced by carcinogenic agents through suppression of IkappaB alpha kinase and p65 phosphorylation: correlation with downregulation of cyclooxygenase 2, matrix metalloproteinase 9, and cyclin D1. Cancer Res. 63, 4375-4383.
  26. Smith, D.W., Wiegeshaus, E., Navakar, R., and Grover, A.A. (1966). Host-parasite relationships in experimental airborne tuberculosis. J. Bacteriol. 91, 718-724.
  27. Stuer, D.J., Gross, S.S., Sukuma, I., Levi, R., and Nathan, C.F. (1989). Activated murine macrophages secrete a metabolite of arginine with the bioactivity of endothelium-derived relaxing factor and the chemical reactivity of nitric oxide. J. Exp. Med. 169, 1011-1020. https://doi.org/10.1084/jem.169.3.1011
  28. Tsai, S.J., and Yin, M.C. (2008). Antioxidative and anti-inflammatory protection of oleanolic acid and ursolic acid in PC12 cells. J. Food Sci. 73, H174-H178. https://doi.org/10.1111/j.1750-3841.2008.00864.x
  29. Xie, Q., and Nathan, C. (1994). The high-output nitric oxide pathway: role and regulation. J. Leukoc. Biol. 56, 576-582. https://doi.org/10.1002/jlb.56.5.576
  30. Yang, J., Hooper, W.C., Phillips, D.J., and Talkington, D.F. (2002). Regulation of proinflammatory cytokines in human lung epithelial cells infected with Mycoplasma pneumoniae. Infect Immun. 70, 3649-3655. https://doi.org/10.1128/IAI.70.7.3649-3655.2002
  31. Yang, J., Hooper, W.C., Phillips, D.J., Tondella, M.L., and Talkington, D.F. (2003). Induction of proinflammatory cytokines in human lung epithelial cells during Chlamydia pneumoniae infection. Infect Immun. 71, 614-620. https://doi.org/10.1128/IAI.71.2.614-620.2003

Cited by

  1. Design, synthesis and in vitro anticancer activity of novel quinoline and oxadiazole derivatives of ursolic acid vol.27, pp.17, 2017, https://doi.org/10.1016/j.bmcl.2017.07.033
  2. Anti-inflammatory potential of ursolic acid in Mycobacterium tuberculosis-sensitized and Concanavalin A-stimulated cells vol.13, pp.3, 2016, https://doi.org/10.3892/mmr.2016.4840
  3. Green Synthesis of Gold Nanoparticles Using Different Leaf Extracts of Ocimum gratissimum Linn for Anti-tubercular Activity vol.9, pp.2, 2015, https://doi.org/10.2174/2468187308666180807125058
  4. Activity of Lactobacillus crispatus isolated from vaginal microbiota against Mycobacterium tuberculosis vol.59, pp.11, 2021, https://doi.org/10.1007/s12275-021-1332-0