Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Raloxifene, a Selective Estrogen Receptor Modulator, Inhibits Lipopolysaccharide-induced Nitric Oxide Production by Inhibiting the Phosphatidylinositol 3-Kinase/Akt/Nuclear Factor-kappa B Pathway in RAW264.7 Macrophage Cells

  • Lee, Sin-Ae (Department of Biochemistry, College of Natural Sciences, Kangwon National University) ;
  • Park, Seok Hee (Department of Biological Science, Sungkyunkwan University) ;
  • Kim, Byung-Chul (Department of Biochemistry, College of Natural Sciences, Kangwon National University)
  • Received : 2007.11.05
  • Accepted : 2007.12.21
  • Published : 2008.07.31
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Abstract

We here demonstrate an anti-inflammatory action of raloxifene, a selective estrogen receptor modulator, in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells. Treatment with raloxifene at micromolar concentrations suppressed the production of nitric oxide (NO) by down-regulating expression of the inducible nitric oxide synthase (iNOS) gene in LPS-activated cells. The decreased expression of iNOS and subsequent reduction of NO were due to inhibition of nuclear translocation of transcription factor NF-${\kappa}B$. These effects were significantly inhibited by exposure to the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY294002, or by expression of a dominant negative mutant of PI 3-kinase. In addition, pretreatment with raloxifene reduced LPS-induced Akt phosphorylation as well as NF-${\kappa}B$ DNA binding activity and NF-${\kappa}B$-dependent reporter gene activity. Thus our findings indicate that raloxifene exerts its anti-inflammatory action in LPS-stimulated macrophages by blocking the PI 3-kinase-Akt-NF-${\kappa}B$ signaling cascade, and eventually reduces expression of pro-inflammatory genes such as iNOS.

Keywords

References

  1. Ariazi, E.A., Ariazi, J.L., Cordera, F., and Jordan, V.C. (2006). Estrogen receptors as therapeutic targets in breast cancer. Curr. Top. Med. Chem. 6, 195-216 https://doi.org/10.2174/156802606776173474
  2. Cirino, G. (1998). Multiple controls in inflammation. Extracellular and intracellular phospholipase A2, inducible and constitutive cyclooxygenase, and inducible nitric oxide synthase. Biochem. Pharmacol. 55, 105-111 https://doi.org/10.1016/S0006-2952(97)00215-3
  3. Delmas, P.D., Bjarnason, N.H., Mitlak, B.H., Ravoux, A.C., Shah, A.S., Huster, W.J., Draper, M., and Christiansen, C. (1997). Effects od raloxifene on bone mineral density, serum cholesterol concentrations, and uterine endometrium in postmenopausal women. N. Engl. J. Med. 337, 1641-1647 https://doi.org/10.1056/NEJM199712043372301
  4. Eming, S.A., Krieg, T., and Davidson, J.M. (2007). Inflammation in wound repair: molecular and cellular mechanisms. J. Invest. Dermatol. 127, 514-525 https://doi.org/10.1038/sj.jid.5700701
  5. Esposito, E., Iacono, A., Raso, G..M., Pacilio, M., Coppola, A., Di Cario, R., and Meli, R. (2005). Raloxifene, a selective estrogen receptor modulator, reduces carrageenan-induced acute inflammation in normal and ovariectomized rats. Endocrinology 146, 3301-3308 https://doi.org/10.1210/en.2005-0375
  6. Grandage, V.L., Gale, R.E., Linch, D.C., and Khwaja, A. (2005). PI3-kinase/Akt is constitutively active in primary acute myeloid leukaemia cells and regulates survival and chemoresistance via NF-kappaB, Mapkinases and p53 pathways. Leukemia 19, 586- 594 https://doi.org/10.1038/sj.leu.2403653
  7. Grese, T.A., Sluka, J.P., Bryant, H.U., Cullinan, G.J., Glasebrook, A.L., Jones, C.D., Matsumoto, K., Palkowitz, A.D., Sato, M., Termine, J.D., et al. (1997). Molecular determinants of tissue selectivity in estrogen receptor modulator. Proc. Natl. Acad. Sci. USA 94, 14105-141107-672
  8. Hanada, T., and Yoshimura, A. (2002). Regulation of cytokine signaling and inflammation. Cytokine Growth Factor Rev. 13, 413- 421 https://doi.org/10.1016/S1359-6101(02)00026-6
  9. Katznellenbogen, B.S., and Katznellenbogen, J.A. (2002). Defining the "S" in SERMS. Science 295, 2380-2381 https://doi.org/10.1126/science.1070442
  10. Kawagoe, J., Ohmichi, M., Takahashi, T., Ohshima, C., Mabuchi, S., Takahashi, K., Igarashi, H., Mori-Abe, A., Saitoh, M., Du, B., et al. (2003). Raloxifene inhibits estrogen-induced up-regulation of telomerase activity in a human breast cancer cell line. J. Biol. Chem. 278, 43363-43372 https://doi.org/10.1074/jbc.M304363200
  11. Kuiper, G.G.J.M., Lemmen, J.G., Carlsson, B., Corton, J.C., Safe, S.H., van der Saag, P.T., vad der Burg, B., and Gustafsson, J.-A. (1998). Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor-$\beta$. Endocrinology 13, 4252-4263
  12. Laskin, D.L., and Pendino, J. (1995). Macrophages and inflammatory mediators in tissue injury. Annu. Rev. Pharmacol. Toxicol. 35, 655-677 https://doi.org/10.1146/annurev.pa.35.040195.003255
  13. Lee, S.J., Bai, S.K., Lee, K.S., Namkoong, S., Na, H.J., Ha, K.S., Han, J.A., Yim, S.Y., Chang, K., Kwon, Y.G., et al. (2003). Astaxanthin inhibits nitric oxide production and inflammatory gene expression by suppressing I(kappa)B kinase-dependent NF-kappaB activation. Mol. Cells 16, 97-105
  14. Lonard, D.M., and Smith, C.L. (2002). Molecular perspectives on selective estrogen receptor modulators (SERM): progress in understanding their tissue-specific agonist antagonist actions. Steroid 67, 15-24 https://doi.org/10.1016/S0039-128X(01)00133-7
  15. Lufkin, E.G., Whitaker, M.D., Nickelsen, T., Argueta, R., Caplan, R.H., Knickerbocker, R.K., and Riggs, B.L. (1998). Treatment of established postmenopausal osteoporosis with raloxifene: A randomized trial. J. Bone Miner. Res. 13, 1747-1754 https://doi.org/10.1359/jbmr.1998.13.11.1747
  16. Romashkova, J.A., and Makarov, S.S. (1999). NF-kappaB is a target of AKT in anti-apoptotic PDGF signaling. Nature 401, 86- 90 https://doi.org/10.1038/43474
  17. Sherman, M.P., Aeberhard, E.E., Wong, V.Z., Griscavage, J.M., and Ignarro, L.J. (1993). Pyrrolidine dithiocarbamate inhibits induction of nitric oxide synthase activity in rat alveolar macrophages. Biochem. Biophys. Res. Commun. 191, 1301-1308 https://doi.org/10.1006/bbrc.1993.1359
  18. Shoji, T., Yoshida, S., Mitsunari, M., Miyake, N., Tsukihara, S., Iwabe, T., Harada, T., and Terakawa, N. (2007). Involvement of p38 MAP kinase in lipopolysaccharide-induced production of pro- and anti-inflammatory cytokines and prostaglandin E(2) in human choriodecidua. J. Reprod. Immunol. 75, 82-90 https://doi.org/10.1016/j.jri.2007.05.002
  19. Stuehr, D.J. (1997). Structure-function aspects in the nitric oxide synthases, Annu. Rev. Pharmacol. Toxicol. 37, 339-359 https://doi.org/10.1146/annurev.pharmtox.37.1.339
  20. Suuronen, T., Nuutinen, T., Huuskonen, J., Ojala, J., Thornell, A., and Salminen, A. (2005). Anti-inflammatory effect of selective estrogen receptor modulators (SERMs) in microglial cells. Inflamm. Res. 54, 194-203 https://doi.org/10.1007/s00011-005-1343-z
  21. Verhaeghe, C., Remouchamps, C., Hennuy, B., Vanderplasschen, A., Chariot, A., Tabruyn, S.P., Oury, C., and Bours, V. (2007). Role of IKK and ERK pathways in intrinsic inflammation of cystic fibrosis airways. Biochem. Pharmacol. 73, 1982-1994 https://doi.org/10.1016/j.bcp.2007.03.019