Molecular and Cellular Biochemistry

, Volume 275, Issue 1–2, pp 165–171 | Cite as

Ganoderma lucidum inhibits inducible nitric oxide synthase expression in macrophages

  • Connie W. H. Woo
  • Ricky Y. K. Man
  • Yaw L. Siow
  • Patrick C. Choy
  • Eric W. Y. Wan
  • Chak S. Lau
  • Karmin O
Article

Abstract

Nitric oxide (NO) is a principal mediator in many physiological and pathological processes. Overproduction of NO via the inducible nitric oxide synthase (iNOS) has cytotoxic effect through the formation of peroxynitrite with superoxide anion. The iNOS is mainly expressed in macrophages and is able to produce large amount of NO. The expression of iNOS is mainly regulated at the transcriptional level. The iNOS-mediated NO production plays a role in the development of atherosclerosis. Ganoderma lucidum (G. lucidum, Linzhi or Reishi) is a traditional herbal medicine which is commonly used as health supplement. Several studies have demonstrated its effectiveness against cancer, immunological disorders and cardiovascular diseases. The objective of the present study was to investigate the effect of G. lucidum on iNOS-mediated NO production in macrophages. Human monocytic cell (THP-1) derived macrophages were incubated with lipopolysaccharide (LPS) for 24 h. Such treatment significantly stimulated NO production (253% versus the control). Such a stimulatory effect was resulted from increased iNOS mRNA expression (270% versus the control) and iNOS activity (169.5% versus the control) in macrophages. The superoxide anion level was also elevated (150% versus the control) in LPS-treated macrophages. Treatment of macrophages with G. lucidum extract (100 μg/ml) completely abolished LPS-induced iNOS mRNA expression and NO production. Such an inhibitory effect of G. lucidum was mediated via its antioxidant action against LPS-induced superoxide anion generation in macrophages. These results suggest that G. lucidum may exert a therapeutic effect against atherosclerosis via ameliorating iNOS-mediated NO overproduction in macrophages.

Keywords

herbal medicine inducible nitric oxide synthase nitric oxide oxidative stress 

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References

  1. 1.
    Shiao M: Natural products of the medicinal fungus Ganoderma lucidum: occurrence, biological activities, and pharmacologial funtions. Chem Rec 3: 172–180, 2003Google Scholar
  2. 2.
    Miyazaki T, Nishijima M: Studies on fungal polysaccharides. XXVII. Structural examination of a water-soluble, antitumor polysaccharide of Ganoderma lucidum. Chem Pharm Bull 71: 115–124, 1981Google Scholar
  3. 3.
    Wang SY, Hsu ML, Hsu HC, Tzeng CH, Lee SS, Shiao MS, Ho CK: The anti-tumor effect of Ganoderma lucidum is mediated by cytokines released from activated marophages and T lymphocytes. Int J Cancer 70: 699–705, 1997Google Scholar
  4. 4.
    Lee SS, Wei YH, Chen CF, Wang SY, Chen KY: Antitumor effects of Ganoderma lucidum. J Chin Med 6: 1–12, 1995Google Scholar
  5. 5.
    Wasser SP, Weis AL: Medicinal properties of substances occurring in higher Basidiomycetes mushrooms: current perspective (review). Int J Med Mushrooms 1: 31–62, 1999Google Scholar
  6. 6.
    Lin S, Li C, Lee S, Kan L: Triterpene-enriched extracts from Ganoderma lucidum inhibit growth of hepatoma cells via suppressing protein kinase C, activating mitogen-activated protein kinases and G2-phase cell cycle arrest. Life Sci 72: 2381–2390, 2003Google Scholar
  7. 7.
    Zhang HN, Lin ZB: Hypoglycemic effect of Gonaderma lucidum polysaccharides. Acta Pharmacol Sin 25(2): 191–195, 2004Google Scholar
  8. 8.
    Hsu MJ, Lee SS, Lee ST, Lin WW: Signaling mechanisms of enhanced neutrophil phagocytosis and chemotaxis by the polysaccharide purified from Ganoderma lucidum. Br J Pharmacol 139(2): 289–298, 2003Google Scholar
  9. 9.
    Zhu M, Chang Q, Wong LK, Chong FS, Li RC: Triterpene antioxidants from Ganoderma lucidum. Phytother Res 13: 529–531, 1999Google Scholar
  10. 10.
    Mau J, Lin H, Chen C: Antioxidant properties of several medicinal mushrooms. J Agric Food Chem 50: 6072–6077, 2002Google Scholar
  11. 11.
    Lee J, Kwon H, Jeong H, Lee JW, Lee SY, Baek SJ, Surh Y: Inhibition of lipid peroxidation and oxidative DNA damage by Ganoderma lucidum. Phytother Res 15: 245–249, 2001Google Scholar
  12. 12.
    Raji L, Baylis C: Glomerular actions of nitric oxide. Kidney Int 48(1): 20–32, 1995Google Scholar
  13. 13.
    Napoli C, Ignarro LJ: Nitric oxide and atherosclerosis. Nitric Oxide 5: 88–97, 2001Google Scholar
  14. 14.
    Radi R, Beckman JS, Bush KM, Freeman BA: Peroxynitrite-induced membrane lipid peroxidation: The cytotoxic potential of superoxide and nitric oxide. Arch Biochem Biophys 288: 481–487, 1991Google Scholar
  15. 15.
    White CR, Brock TA, Chang LT, et al.: Superoxide and peroxynitrite in atherosclerosis. Proc Natl Acad Sci USA 91: 1044–1048, 1994Google Scholar
  16. 16.
    Estévez A, Jordán J: Nitric oxide and superoxide, a deadly cocktail. Ann N Y Acad Sci 962: 207–211, 2002Google Scholar
  17. 17.
    Schussheim AE, Fuster V: Antibiotics for myocardial infarction? A possible role of infection in atherogenesis and acute coronary syndromes. Drug 57(3): 283–291, 1999Google Scholar
  18. 18.
    Rueckschloss U, Duerrschmidt N, Morawietz H: NADPH oxidase in endothelial cells: Impact on atherosclerosis. Antioxid Redox Signal 5(2): 171–180, 2003Google Scholar
  19. 19.
    Liu S, Adcock IM, Old RW, Barnes PJ, Evans TW: Lipopolysaccharide treatment in vivo induces widespread tissue expression of inducible nitric oxide synthase mRNA. Biochem Biophys Res Commun 196: 1208–1213, 1993Google Scholar
  20. 20.
    Cheung F, Siow YL, Karmin O: Inhibition by ginkgolides and bilobalide of the production of nitric oxide in macrophages (THP-1) but not in endothelial cells (HUVEC). Biochem Pharmacol 61: 503–510, 2001Google Scholar
  21. 21.
    Wang G, Siow YL, Karmin O: Homocysteine induces monocyte chemoattractant protein-1 expression by activating NF-kappa B in THP-1 macrophage. Am J Physiol Heart Cir Physiol 280: H2840–H2847, 2001Google Scholar
  22. 22.
    Kurosaka K, Watanabe N, Kabayashi Y: Production of proinflammatory cytokines by phorbol myristate acetate-treated THP-1 cells and monocyte-derived macrophages after phagocytosis of apoptotic CTLL-2 cells. J Immunol 161: 6245–6249, 1998Google Scholar
  23. 23.
    Wang N, Tabas I, Winchester R, Ravalli S, Rabbani LE, Tall A: Interleukin 8 is induced by cholesterol loading of macrophage and expressed by macrophage foam cells in human atheroma. J Biol Chem 271: 8837–8842, 1996Google Scholar
  24. 24.
    Michelakis ED, Archer SL: The measurement of NO in biological systems using chemiluminescene. In: M. Titheradge (ed.). Nitric Oxide Protocols, 1998, pp 111–127Google Scholar
  25. 25.
    Schmidt H, Kelm M: Determination of nitrite and nitrate by the griess reaction. In: M. Feelisch, J.S. Stamler (eds.). Methods in Nitric Oxide Research, 1996, pp 491–497Google Scholar
  26. 26.
    Bredti DS, Shmidt H: The Citrulline assay. In: M. Feelisch, J.S. Stamler (eds.). Methods in Nitric Oxide Research. J. Wiley: Chichester, New York, 1996, pp 249–255Google Scholar
  27. 27.
    Wang G, Siow YL, Karmin O: Homocysteine stimulates nuclear factor kappa B activity and monocyte chemoattractant protein-1 expression in vascular smooth muscle cells: a possible role for protein kinase C. Biochem J 352: 817–826, 2000Google Scholar
  28. 28.
    Rauen U, Petrat F, Li T, De Groot H: Hypothermia injury/cold-induced apoptosis – evidence of an increase in chelatable iron causing oxidative injury in spite of low O2/H2O2 formation. FASEB J 14: 1953–1964, 2000Google Scholar
  29. 29.
    Woo CW, Cheung F, Chan VW, Siow YL, Karmin O: Homocysteine stimulates inducible nitric oxide synthase expression in macrophages: antagonizing effect of ginkgolides and bilobalide. Mol Cell Biochem 243(1–2): 37–47, 2003Google Scholar
  30. 30.
    Komoda Y, Shimizu M, Sonoda Y, Sato Y: Ganoderic acid and its derivatives as cholesterol synthesis inhibitors. Chem Pharm Bull (Tokyo) 37(2): 531–533, 1989Google Scholar
  31. 31.
    Kabir Y, Kimura S, Tamura T: Dietary effect of Ganoderma lucidum mushroom on blood pressure and lipid levels in spontaneously hypertensive rats (SHR). J Nutr Sci Vitaminol (Tokyo) 34(4): 433–438, 1998Google Scholar
  32. 32.
    Tao J, Feng KY: Experimental and clinical studies on inhibitory effect of Ganoderma lucidum on platelet aggregation. J Tongji Med Univ 10(4): 240–243, 1990Google Scholar
  33. 33.
    Khatri JJ, Johnson C, Magid R, Lessner SM, Laude KM, Dikalov SI, Harrison DG, Sung HJ, Rong Y, Galis ZS: Vascular oxidant stress enhances progression and angiogenesis of experimental atheroma. Circulation 109: 520–525, 2004Google Scholar
  34. 34.
    Juliet PA, Hayashi T, Daigo S, Matsui-Hirai H, Miyazaki A, Fukatsu A, Funami J, Iguchi A, Ignarro LJ: Combined effect of testosterone and apocynin on nitric oxide and superoxide production in PMA-differentiated THP-1 cells. Biochim Biophys Acta 1693(3): 185–191, 2004Google Scholar
  35. 35.
    Begarja AA, Wu D, Serafini M, Meydani SN: Mechanism of vitamin E inhibition of cyclooxygenase activity in macrophages from old mice: role of peroxynitrite. Free Radic Biol Med 32(6): 503–511, 2002Google Scholar
  36. 36.
    Lancel S, Tissier S, Mordon S, Marechal X, Depontieu F, Scherpereel A, Chopin C, Neviere R: Peroxynitrite decomposition catalysts prevent myocardial dysfunction and inflammation in endotoxemic rats. J Am Coll Cardiol 43(12): 2348–2358, 2004Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Connie W. H. Woo
    • 1
  • Ricky Y. K. Man
    • 2
  • Yaw L. Siow
    • 1
    • 3
    • 4
  • Patrick C. Choy
    • 4
  • Eric W. Y. Wan
    • 2
  • Chak S. Lau
    • 5
  • Karmin O
    • 1
    • 3
    • 6
    • 7
  1. 1.Department of PhysiologyUniversity of ManitobaWinnipegCanada
  2. 2.National Centre for Agri-Food Research in Medicine, St. Boniface Hospital Research CentreUniversity of ManitobaWinnipegCanada
  3. 3.Department of PharmacologyUniversity of Hong KongCanada
  4. 4.Centre for Research and Treatment of AtherosclerosisUniversity of ManitobaWinnipegCanada
  5. 5.Department of MedicineUniversity of Hong KongCanada
  6. 6.Department of Animal ScienceUniversity of ManitobaWinnipegCanada
  7. 7.Laboratory of Integrative BiologySt. Boniface Hospital Research CentreWinnipegCanada

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