Molecular and Cellular Biochemistry

, Volume 276, Issue 1–2, pp 7–13 | Cite as

Inhibition of radiation induced nitration by curcumin and nicotinamide in mouse macrophages

  • Himanshi Narang
  • Malini Krishna


Nitric oxide plays an important role in inflammation and carcinogenesis and has now been implicated as an important signaling molecule under normal physiological conditions also. Increased nitric oxide (NO) results in increased nitration of proteins at tyrosine, which can cause protein dysfunction or alterations in signal transduction pathways. Irradiation of Lipopolysaccharide (LPS) activated mouse peritoneal macrophages was found to increase NO production, inducible nitric oxide synthase (iNOS) expression and nitration of proteins. The increase in iNOS expression was very less when compared to increase in NO production, indicating the possibility of post-translational activation of iNOS by LPS and ionising radiation. The addition of curcumin, nicotinamide and Jun N-terminal kinase (JNK) inhibitor, SP600125, reduced the levels of NO, iNOS expression and nitration of proteins in macrophages. Closer scrutiny of the inhibition pattern of these modulators revealed that although the JNK inhibitor did not result in significant decrease in iNOS expression it led to a significant decrease in NO production, implying the possible involvement of JNK in the regulation of iNOS activity. Curcumin and JNK inhibitor directly inhibited the nitration of proteins and JNK inhibitor and curcumin, when added together, did not show synergistic effect. (Mol Cell Biochem 276: 7–13, 2005)


curcumin iNOS irradiation JNK inhibitor II macrophages nicotinamide tyrosine nitration 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Nathan C, Xie QW: Regulation of biosynthesis of nitric oxide. J Biol Chem 269: 13725–13728, 1994PubMedGoogle Scholar
  2. 2.
    Lowenstein CJ, Snyder SH: Nitric oxide, a novel biological messenger. Cell 70: 705–707, 1992CrossRefPubMedGoogle Scholar
  3. 3.
    Ibuki Y, Goto R: Enhancement of NO production from resident peritoneal macrophages by in vitro γ-irradiation and its relationships to reactive oxygen intermediates. Free Radic Biol Med 22: 1029–1035, 1997CrossRefPubMedGoogle Scholar
  4. 4.
    Ibuki Y, Mizuno S, Goto R: γ-irradiation induced DNA damage enhances NO production via NF-κB activation in RAW264.7 cells. Biochim Biophys Acta 1593: 159–167, 2003CrossRefPubMedGoogle Scholar
  5. 5.
    Jun C-D, Oh C-D, Kwak H-J,Pae H-O, Yoo J-C, Choi B-M, Chun J-S, Park R-K, Chung H-T: Overexpression of protein kinase C isoforms protects RAW 264.7 macrophages from nitric oxide induced apoptosis: involvement of c-jun N-terminal kinase/stress activated kinase, p38 kinase, and CPP-32 protease pathways. J Immunol 162: 3395–3401, 1999PubMedGoogle Scholar
  6. 6.
    St-Denis A, Chano F, Tremblay P, St-Pierre Y, Decoteaux A: Protein kinase C-alpha modulates lipopolysaccharide induced functions in murine macrophage cell line. J Biol Chem 273: 32787–32792, 1998CrossRefPubMedGoogle Scholar
  7. 7.
    Chan ED, Riches DW: IFN-γ + LPS induction of iNOS is modulated byERK, JNK/SAPK and p38 (MAPK) in mouse macrophage cell line. Am J Physiol Cell Physiol 280: C441–450, 2001PubMedGoogle Scholar
  8. 8.
    Chan ED, Winston BW, Uh ST, Wynes MW, Rose DM, Riches DWH: Evaluation of the role of mitogen activated protein kinases in the expression of inducible nitric oxide synthase by IFN-γ and TNF-γ in mouse macrophages. J Immunol 162: 415–422, 1999PubMedGoogle Scholar
  9. 9.
    Inoue S, Kawanishi S: Oxidative damage induced by simultaneous generation of nitric oxide and superoxide. FEBS Lett 371: 86–88, 1995CrossRefPubMedGoogle Scholar
  10. 10.
    Ischiropoulos H, Al-Mehdi AB: Peroxynitrite-mediated oxidative protein modifications. FEBS Lett 364: 279–282, 1995CrossRefPubMedGoogle Scholar
  11. 11.
    Ischiropoulos H, Zhu L, Chen J, Tsai M, Martin JC, Smith CD, Beckman JS: Peroxynitrite mediated tyrosine nitration catalyzed by superoxide dismutase. Arch Biochem Biophys 298: 431–437, 1992CrossRefPubMedGoogle Scholar
  12. 12.
    Greenacre SAB, Ischiropoulos H: Tyrosine nitration: Localisation, quantification, consequences for protein function and signal transduction. Free Rad Res 34: 541–581, 2001Google Scholar
  13. 13.
    Ischiropoulos H: Biological tyrosine nitration: A pathophysiological function of nitric oxide and reactive oxygen species. Arch Biochem Biophys 356: 1–11, 1998CrossRefPubMedGoogle Scholar
  14. 14.
    Knapp LT, Kanterwicz BI, Hayes EL, Klann E: Peroxynitrite-induced tyrosine nitration and inhibition of protein kinase C. Biochem Biophys Res Commun 286: 764–770, 2001CrossRefPubMedGoogle Scholar
  15. 15.
    Brito C, Naviliat M, Tiscornia AC, Vuillier F, Gualco G, Dighiero G, Radi R, Cayota AM: Peroxynitrite inhibits T lymphocyte activation and proliferation by promoting impairment of tyrosine phosphorylation and peroxynitrite driven apoptotic death. J Immunol 162: 3356–3366, 1999PubMedGoogle Scholar
  16. 16.
    Monteiro HP: Signal transduction by protein tyrosine nitration: Competition or cooperation with tyrosine phosphorylation dependent signaling events. Free Radical Biol Med 33 (6): 765–773, 2002CrossRefGoogle Scholar
  17. 17.
    Lepoivre M, Fieschi F, Coves J, Thelander J, Fontecave M: Inactivation of ribonucleotide reductase by nitric oxide. Biochem Biophys Res Commun 179: 422–428, 1991CrossRefGoogle Scholar
  18. 18.
    Cassina A, Radi R: Differential inhibitory action of nitric oxide and peroxynitrite on mitochondrial electron transport. Arch Biochem Biophys 328: 309–316, 1996CrossRefPubMedGoogle Scholar
  19. 19.
    MacMicking J, Xie QW, Nathan C: Nitic oxide and macrophage function. Annu Rev Immunol 15: 323–350, 1997CrossRefPubMedGoogle Scholar
  20. 20.
    Sandoval M, Zhang XJ, Liu X, Mannick EE, Clark DA, Miller MJ. Peroxynitrite induced apoptosis in T84 and RAW 264.7 cells: Attenuation by L-Ascorbic acid. Free Radic Biol Med 22: 489–495, 1997CrossRefPubMedGoogle Scholar
  21. 21.
    Liu JY, Lin SJ, Lin JK: Inhibitory effects of curcumin on protein kinase C activity induced by 12-O-tetradecanoylphorbol-13-acetate in NIH-3T3 cells. Carcinogenesis 14: 857–861, 1993PubMedGoogle Scholar
  22. 22.
    Varadkar P, Dubey P, Krishna M, Verma NC: Modulation of radiation-induced protein kinase C activity by phenolics. J Radiol Prot 21: 361–370, 2001CrossRefPubMedGoogle Scholar
  23. 23.
    Mitra AK, Krishna M: In vivo modulation of signaling factors involved in cell survival. J Radiat Res 45(4): 491–495, 2004CrossRefPubMedGoogle Scholar
  24. 24.
    Choudhury S, Krishna M, Bhattacharya RK: Modulation of NDEA activated ras expression and protein kinase C activity by nicotinamide. Cancer Lett 147: 39–44, 1999CrossRefPubMedGoogle Scholar
  25. 25.
    Brouet I, Ohshima H: Curcumin, an anti-tumor promoter and anti-inflammatory agent, inhibits induction of nitric oxide synthase in activated macrophages. Biochem Biophys Res Commun 206: 535–540, 1995CrossRefGoogle Scholar
  26. 26.
    Chan MMY, Huang HI, Fenton MR, Fong D: In vivo inhibition of nitric oxide synthase gene expression by curcumin, a cancer preventive natural product with anti-inflammatory properties. Biochem Pharmacol 55: 1955–1962, 1998CrossRefPubMedGoogle Scholar
  27. 27.
    Pan MH, Lin-Shiau S, Lin JK: Comparative studies on the suppression of nitric oxide synthase by curcumin and its hydrogenated metabolites through down regulation of IκB Kinase and NF-κB activation in macrophages. Biochem Pharmacol 60: 1665–1676, 2000CrossRefPubMedGoogle Scholar
  28. 28.
    Le Page C, Sanceau J, Drapier JC, Wietzerbin J: Inhibitors of ADP-ribosylation impair inducible nitric oxide synthase gene transcription through inhibition of NF-κB activation. Biochem Biophys Res Commun 243: 451–457, 1998CrossRefPubMedGoogle Scholar
  29. 29.
    Bhaumik S, Jyothi MD, Khar A: Differential modulation of nitric oxide production by curcumin in host macrophages and NK cells. FEBS Lett 483: 78–82, 2000CrossRefPubMedGoogle Scholar
  30. 30.
    Ibuki Y, Goto R: Enhancement of concanavalin A-induced proliferation of spleno-lymphocytes by low-dose-irradiated macrophages. J Radiat Res 35: 83–91, 1994PubMedGoogle Scholar
  31. 31.
    Schieke SM, Briviba K, Klotz LO, Sies H: Activation pattern of mitogen-activated protein kinases elicited by peroxynitrite: Attenuation by selenite supplementation. FEBS Lett 448: 301–303, 1999CrossRefPubMedGoogle Scholar
  32. 32.
    Johnston BD, DeMaster EG: Suppresion of nitric oxide oxidation to nitrite by curcumin is due to sequestration of the reaction intermediate nitrogen dioxide, not nitric oxide. Nitric Oxide 8: 231–234, 2003CrossRefPubMedGoogle Scholar
  33. 33.
    Bhat NR, Zhang P, Lee JC, Hogan EL: Extracellular signal-regulated kinase and p38 subgroups of mitogen-activated protein kinases regulate inducible nitric oxide synthase and tumor necrosis factor-κ gene expression in endotoxin-stimulated primary glial cultures. J Neurosci 18: 1633–1637, 1998PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Radiation Biology and Health Sciences DivisionBhabha Atomic Research CentreMumbaiIndia
  2. 2.Radiation Biology and Health Sciences DivisionB. A. R. C.MumbaiIndia

Personalised recommendations