Skip to main content
SpringerLink
Log in

Ketamine Inhibits Calcium Elevation and Hydroxyl Radical and Nitric Oxide Production in Lipopolysaccharide-Stimulated NR8383 Alveolar Macrophages

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Macrophages play a critical role in mediating inflammatory processes; activated macrophages respond to endotoxin by releasing pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6. Ketamine, a widely used anesthetic agent, has unequivocally anti-inflammatory effects in vivo and in vitro. However, the detailed mechanisms for the anti-inflammatory effects of ketamine in microglia have not been elucidated yet. This study aimed to evaluate the effects of ketamine on lipopolysaccharide (LPS)-induced nitric oxide (NO), hydroxyl radical (·OH) production, and intracellular calcium accumulation in macrophages. Macrophages were pretreated with ketamine at the concentrations of 10, 100, and 1,000 μM 1 h before LPS stimulation. The production of NO and ·OH in the culture supernatant of macrophages was assayed by Griess Reagent Kit. LPS enhanced NO and ·OH production and provoked a significant intracellular calcium elevation. With the concentrations higher than 100 μM, ketamine inhibited LPS-induced NO and ·OH accumulation and intracellular calcium elevation. However, a low concentration of ketamine (10 μM) did not exert anti-inflammatory effects. These results suggest that intracellular calcium elevation is, at least, partially involved in the inhibitory effect of ketamine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Doherty, G.M., J.R. Lange, H.N. Langstein, H.R. Alexander, C.M. Buresh, and J.A. Norton. 1992. Evidence for IFN-gamma as a mediator of the lethality of endotoxin and tumor necrosis factor-alpha. Journal of Immunology 149: 1666–1670.

    CAS  Google Scholar 

  2. Nakagawa, S., Y. Arai, T. Kishida, N. Hiraoka, S. Tsuchida, H. Inoue, R. Sakai, O. Mazda, and T. Kubo. 2012. Lansoprazole inhibits nitric oxide and prostaglandin E(2) production in murine macrophage RAW 264.7 cells. Inflammation 35: 1062–1068.

    Article  PubMed  CAS  Google Scholar 

  3. Valledor, A.F., and M. Ricote. 2004. Nuclear receptor signaling in macrophages. Biochemical Pharmacology 67: 201–212.

    Article  PubMed  CAS  Google Scholar 

  4. Nathan, C.F. 1987. Neutrophil activation on biological surfaces. Massive secretion of hydrogen peroxide in response to products of macrophages and lymphocytes. The Journal of Clinical Investigation 80: 1550–1560.

    Article  PubMed  CAS  Google Scholar 

  5. Wu, G.J., T.L. Chen, Y.F. Ueng, and R.M. Chen. 2008. Ketamine inhibits tumor necrosis factor-alpha and interleukin-6 gene expressions in lipopolysaccharide-stimulated macrophages through suppression of toll-like receptor 4-mediated c-Jun N-terminal kinase phosphorylation and activator protein-1 activation. Toxicology and Applied Pharmacology 228: 105–113.

    Article  PubMed  CAS  Google Scholar 

  6. Shimaoka, M., T. Iida, A. Ohara, N. Taenaka, T. Mashimo, T. Honda, and I. Yoshiya. 1996. Ketamine inhibits nitric oxide production in mouse-activated macrophage-like cells. British Journal of Anaesthesia 77: 238–242.

    Article  PubMed  CAS  Google Scholar 

  7. Gill, R., A. Tsung, and T. Billiar. 2010. Linking oxidative stress to inflammation: Toll-like receptors. Free Radical Biology & Medicine 48: 1121–1132.

    Article  CAS  Google Scholar 

  8. Chang, Y., T.L. Chen, J.R. Sheu, and R.M. Chen. 2005. Suppressive effects of ketamine on macrophage functions. Toxicology and Applied Pharmacology 204: 27–35.

    Article  PubMed  CAS  Google Scholar 

  9. Sato, S., O. Takeuchi, T. Fujita, H. Tomizawa, K. Takeda, and S. Akira. 2002. A variety of microbial components induce tolerance to lipopolysaccharide by differentially affecting MyD88-dependent and -independent pathways. International Immunology 14: 783–791.

    Article  PubMed  CAS  Google Scholar 

  10. Reddy, D.B., and P. Reddanna. 2009. Chebulagic acid (CA) attenuates LPS-induced inflammation by suppressing NF-kappaB and MAPK activation in RAW 264.7 macrophages. Biochemical and Biophysical Research Communications 381: 112–117.

    Article  PubMed  CAS  Google Scholar 

  11. Gentile, C., M. Allegra, F. Angileri, A.M. Pintaudi, M.A. Livrea, and L. Tesoriere. 2012. Polymeric proanthocyanidins from Sicilian pistachio (Pistacia vera L.) nut extract inhibit lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. European Journal of Nutrition 51: 353–363.

    Article  PubMed  CAS  Google Scholar 

  12. Sun, J., Z.Q. Zhou, R. Lv, W.Y. Li, and J.G. Xu. 2004. Ketamine inhibits LPS-induced calcium elevation and NF-kappa B activation in monocytes. Inflammation Research 53: 304–308.

    Article  PubMed  CAS  Google Scholar 

  13. White, P.F., W.L. Way, and A.J. Trevor. 1982. Ketamine—its pharmacology and therapeutic uses. Anesthesiology 56: 119–136.

    Article  PubMed  CAS  Google Scholar 

  14. Bourgoin, A., J. Albanese, N. Wereszczynski, M. Charbit, R. Vialet, and C. Martin. 2003. Safety of sedation with ketamine in severe head injury patients: comparison with sufentanil. Critical Care Medicine 31: 711–717.

    Article  PubMed  CAS  Google Scholar 

  15. Chen, T.L., C.C. Chang, Y.L. Lin, Y.F. Ueng, and R.M. Chen. 2009. Signal-transducing mechanisms of ketamine-caused inhibition of interleukin-1 beta gene expression in lipopolysaccharide-stimulated murine macrophage-like Raw 264.7 cells. Toxicology and Applied Pharmacology 240: 15–25.

    Article  PubMed  CAS  Google Scholar 

  16. Chang, Y., J.J. Lee, C.Y. Hsieh, G. Hsiao, D.S. Chou, and J.R. Sheu. 2009. Inhibitory effects of ketamine on lipopolysaccharide-induced microglial activation. Mediators of Inflammation 2009: 705379.

    Article  PubMed  Google Scholar 

  17. Kawasaki, T., M. Ogata, C. Kawasaki, J. Ogata, Y. Inoue, and A. Shigematsu. 1999. Ketamine suppresses proinflammatory cytokine production in human whole blood in vitro. Anesthesia and Analgesia 89: 665–669.

    PubMed  CAS  Google Scholar 

  18. Jiang, W., and D.S. Pisetsky. 2006. The role of IFN-alpha and nitric oxide in the release of HMGB1 by RAW 264.7 cells stimulated with polyinosinic-polycytidylic acid or lipopolysaccharide. Journal of Immunology 177: 3337–3343.

    CAS  Google Scholar 

  19. Chang, H.C., K.H. Lin, Y.T. Tai, J.T. Chen, and R.M. Chen. 2010. Lipoteichoic acid-induced TNF-alpha and IL-6 gene expressions and oxidative stress production in macrophages are suppressed by ketamine through downregulating Toll-like receptor 2-mediated activation oF ERK1/2 and NFkappaB. Shock 33: 485–492.

    Article  PubMed  CAS  Google Scholar 

  20. Scorrano, L., S.A. Oakes, J.T. Opferman, E.H. Cheng, M.D. Sorcinelli, T. Pozzan, and S.J. Korsmeyer. 2003. BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 300: 135–139.

    Article  PubMed  CAS  Google Scholar 

  21. Przygodzki, T., A. Sokal, and M. Bryszewska. 2005. Calcium ionophore A23187 action on cardiac myocytes is accompanied by enhanced production of reactive oxygen species. Biochimica et Biophysica Acta 1740: 481–488.

    Article  PubMed  CAS  Google Scholar 

  22. Jou, M.J., T.I. Peng, L.F. Hsu, S.B. Jou, R.J. Reiter, C.M. Yang, C.C. Chiao, Y.F. Lin, and C.C. Chen. 2010. Visualization of melatonin's multiple mitochondrial levels of protection against mitochondrial Ca(2+)-mediated permeability transition and beyond in rat brain astrocytes. Journal of Pineal Research 48: 20–38.

    Article  PubMed  CAS  Google Scholar 

  23. Petersen, A., R.F. Castilho, O. Hansson, T. Wieloch, and P. Brundin. 2000. Oxidative stress, mitochondrial permeability transition and activation of caspases in calcium ionophore A23187-induced death of cultured striatal neurons. Brain Research 857: 20–29.

    Article  PubMed  CAS  Google Scholar 

  24. Spentzas, T., R. Kudumula, C. Acuna, A.J. Talati, K.C. Ingram, F. Savorgnan, E.A. Meals, and B.K. English. 2011. Role of bacterial components in macrophage activation by the LAC and MW2 strains of community-associated, methicillin-resistant Staphylococcus aureus. Cellular Immunology 269: 46–53.

    Article  PubMed  CAS  Google Scholar 

  25. Spentzas, T., R.K. Shapley, C.A. Aguirre, E. Meals, L. Lazar, M.S. Rayburn, B.S. Walker, and B.K. English. 2011. Ketamine inhibits tumor necrosis factor secretion by RAW264.7 murine macrophages stimulated with antibiotic-exposed strains of community-associated, methicillin-resistant Staphylococcus aureus. BMC Immunol 12: 11.

    Article  PubMed  CAS  Google Scholar 

  26. Helmke, R.J., V.F. German, and J.A. Mangos. 1989. A continuous alveolar macrophage cell line: comparisons with freshly derived alveolar macrophages. In Vitro Cellular & Developmental Biology: Journal of the Tissue Culture Association 25: 44–48.

    Article  CAS  Google Scholar 

  27. Aldakkak, M., D.F. Stowe, R.K. Dash, and A.K. Camara. 2012. Mitochondrial handling of excess Ca(2+) is substrate-dependent with implications for reactive oxygen species generation. Free Radical Biology & Medicine. doi:10.1016/j.freeradbiomed.2012.09.020.

    Google Scholar 

  28. Stowe, D.F., and A.K. Camara. 2009. Mitochondrial reactive oxygen species production in excitable cells: modulators of mitochondrial and cell function. Antioxidants & Redox Signaling 11: 1373–1414.

    Article  CAS  Google Scholar 

  29. Tsutsui, H., S. Kinugawa, and S. Matsushima. 2008. Oxidative stress and mitochondrial DNA damage in heart failure. Circulation Journal 72(Suppl A): A31–37.

    Article  PubMed  Google Scholar 

  30. D'Acquisto, F., M.J. May, and S. Ghosh. 2002. Inhibition of nuclear factor kappa B (NF-B): an emerging theme in anti-inflammatory therapies. Molecular Interventions 2: 22–35.

    Article  PubMed  Google Scholar 

  31. Yu, Y., Z. Zhou, J. Xu, Z. Liu, and Y. Wang. 2002. Ketamine reduces NFkappaB activation and TNFalpha production in rat mononuclear cells induced by lipopolysaccharide in vitro. Annals of Clinical and Laboratory Science 32: 292–298.

    PubMed  CAS  Google Scholar 

  32. Yildiz, H., I. Coskuner, E. Bulbuloglu, E. Silay, E.B. Kurutas, Z. Dogan, B. Kantarceken, H. Oksuz, N. Senoglu, M.F. Yuzbasioglu, A. Cetinkaya, and H. Ciralik. 2012. The protective effects of ketamine and propofol in obstructive jaundice: an experimental study. Bratislavské Lekárske Listy 113: 139–144.

    PubMed  CAS  Google Scholar 

  33. Hou, L., and X. Wang. 2001. PKC and PKA, but not PKG mediate LPS-induced CGRP release and [Ca(2+)](i) elevation in DRG neurons of neonatal rats. Journal of Neuroscience Research 66: 592–600.

    Article  PubMed  CAS  Google Scholar 

  34. Jou, M.J., T.I. Peng, H.Y. Wu, and Y.H. Wei. 2005. Enhanced generation of mitochondrial reactive oxygen species in cybrids containing 4977-bp mitochondrial DNA deletion. Annals of the New York Academy of Sciences 1042: 221–228.

    Article  PubMed  CAS  Google Scholar 

  35. Peng, T.I., P.R. Yu, J.Y. Chen, H.L. Wang, H.Y. Wu, Y.H. Wei, and M.J. Jou. 2006. Visualizing common deletion of mitochondrial DNA-augmented mitochondrial reactive oxygen species generation and apoptosis upon oxidative stress. Biochimica et Biophysica Acta 1762: 241–255.

    Article  PubMed  CAS  Google Scholar 

  36. Jou, M.J., T.I. Peng, P.Z. Yu, S.B. Jou, R.J. Reiter, J.Y. Chen, H.Y. Wu, C.C. Chen, and L.F. Hsu. 2007. Melatonin protects against common deletion of mitochondrial DNA-augmented mitochondrial oxidative stress and apoptosis. Journal of Pineal Research 43: 389–403.

    Article  PubMed  CAS  Google Scholar 

  37. Grijalba, M.T., A.E. Vercesi, and S. Schreier. 1999. Ca2+−induced increased lipid packing and domain formation in submitochondrial particles. A possible early step in the mechanism of Ca2+−stimulated generation of reactive oxygen species by the respiratory chain. Biochemistry 38: 13279–13287.

    Article  PubMed  CAS  Google Scholar 

  38. Duan, Y., R.A. Gross, and S.S. Sheu. 2007. Ca2+−dependent generation of mitochondrial reactive oxygen species serves as a signal for poly(ADP-ribose) polymerase-1 activation during glutamate excitotoxicity. The Journal of Physiology 585: 741–758.

    Article  PubMed  CAS  Google Scholar 

  39. Li, C.Y., T.C. Chou, C.S. Wong, S.T. Ho, C.C. Wu, M.H. Yen, and Y.A. Ding. 1997. Ketamine inhibits nitric oxide synthase in lipopolysaccharide-treated rat alveolar macrophages. Canadian Journal of Anaesthesia 44: 989–995.

    Article  PubMed  CAS  Google Scholar 

  40. Weigand, M.A., H. Schmidt, Q. Zhao, K. Plaschke, E. Martin, and H.J. Bardenheuer. 2000. Ketamine modulates the stimulated adhesion molecule expression on human neutrophils in vitro. Anesthesia and Analgesia 90: 206–212.

    Article  PubMed  CAS  Google Scholar 

  41. Zhang, J., B. Jin, L. Li, E.R. Block, and J.M. Patel. 2005. Nitric oxide-induced persistent inhibition and nitrosylation of active site cysteine residues of mitochondrial cytochrome-c oxidase in lung endothelial cells. American Journal of Physiology. Cell Physiology 288: C840–849.

    Article  PubMed  CAS  Google Scholar 

  42. Peng, T.I., and M.J. Jou. 2010. Oxidative stress caused by mitochondrial calcium overload. Annals of the New York Academy of Sciences 1201: 183–188.

    Article  PubMed  CAS  Google Scholar 

  43. Grant, I.S., W.S. Nimmo, L.R. McNicol, and J.A. Clements. 1983. Ketamine disposition in children and adults. British Journal of Anaesthesia 55: 1107–1111.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anesthesiology, The First People’s Hospital of Lianyungang City, Lianyungang, Jiangsu Province, People’s Republic of China

    Xiaobao Zhang, Jiying Feng, Pin Zhu & Zhibin Zhao

Authors
  1. Xiaobao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jiying Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pin Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhibin Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhibin Zhao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, X., Feng, J., Zhu, P. et al. Ketamine Inhibits Calcium Elevation and Hydroxyl Radical and Nitric Oxide Production in Lipopolysaccharide-Stimulated NR8383 Alveolar Macrophages. Inflammation 36, 1094–1100 (2013). https://doi.org/10.1007/s10753-013-9642-y

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-013-9642-y

KEY WORDS

Search

Navigation