Molecular Medicine

, Volume 14, Issue 7–8, pp 422–428 | Cite as

Lysophosphatidic Acid Inhibits Bacterial Endotoxin-Induced Pro-Inflammatory Response: Potential Anti-Inflammatory Signaling Pathways

  • Hongkuan Fan
  • Basilia Zingarelli
  • Vashaunta Harris
  • George E. Tempel
  • Perry V. Halushka
  • James A. Cook
Research Article


Previous studies have demonstrated that heterotrimeric guanine nucleotide-binding regulatory (Gi) protein-deficient mice exhibit augmented inflammatory responses to lipopolysaccharide (LPS). These findings suggest that Gi protein agonists will suppress LPS-induced inflammatory gene expression. Lysophosphatidic acid (LPA) activates G protein-coupled receptors leading to Gi protein activation. We hypothesized that LPA will inhibit LPS-induced inflammatory responses through activation of Gi-coupled anti-inflammatory signaling pathways. We examined the anti-inflammatory effect of LPA on LPS responses both in vivo and in vitro in CD-1 mice. The mice were injected intravenously with LPA (10 mg/kg) followed by intraperitoneal injection of LPS (75 mg/kg for survival and 25 mg/kg for other studies). LPA significantly increased the mice survival to endotoxemia (P < 0.05). LPA injection reduced LPS-induced plasma TNF-α production (69 ± 6%, P < 0.05) and myeloperoxidase (MPO) activity in lung (33 ± 9%, P < 0.05) as compared to vehicle injection. LPS-induced plasma IL-6 was unchanged by LPA. In vitro studies with peritoneal macrophages paralleled results from in vivo studies. LPA (1 and 10 µM) significantly inhibited LPS-induced TNFα production (61 ± 9% and 72 ± 9%, respectively, P < 0.05) but not IL-6. We further demonstrated that the anti-inflammatory effect of LPA was reversed by ERK 1/2 and phosphatase inhibitors, suggesting that ERK 1/2 pathway and serine/threonine phosphatases are involved. Inhibition of phosphatidylinositol 3 (PI3) kinase signaling pathways also partially reversed the LPA anti-inflammatory response. However, LPA did not alter NFκB and peroxisome proliferator-activated receptor γ (PPARγ) activation. Inhibitors of PPARγ did not alter LPA-induced inhibition of LPS signaling. These studies demonstrate that LPA has significant anti-inflammatory activities involving activation of ERK 1/2, serine/threonine phosphatases, and PI3 kinase signaling pathways.



This work was supported by NIH GM27673 and NIH GM67202.


  1. 1.
    Janeway CA Jr, Medzhitov R. (1999) Lipoproteins take their toll on the host. Curr. Biol. 9:R879–82.CrossRefGoogle Scholar
  2. 2.
    Medzhitov R, Janeway CA Jr. (1997) Innate immunity: impact on the adaptive immune response. Curr. Opin. Immunol. 9:4–9.CrossRefGoogle Scholar
  3. 3.
    Glauser MP, Zanetti G, Baumgartner JD, Cohen J. (1991) Septic shock: pathogenesis. Lancet. 338:732–6.CrossRefGoogle Scholar
  4. 4.
    Fan H, Peck OM, Tempel GE, Halushka PV, Cook JA. (2004) Toll-like receptor 4 coupled GI protein signaling pathways regulate extracellular signalregulated kinase phosphorylation and AP-1 activation independent of NF kappa B activation. Shock. 22:57–62.CrossRefGoogle Scholar
  5. 5.
    Fan H et al. (2006) Differential regulation of lipopolysaccharide and Gram-positive bacteria induced cytokine and chemokine production in splenocytes by Galphai proteins. Biochim. Biophys. Acta 1763:1051–8.CrossRefGoogle Scholar
  6. 6.
    Fan H et al. (2005) Lipopolysaccharide- and grampositive bacteria-induced cellular inflammatory responses: role of heterotrimeric Galpha(i) proteins. Am. J. Physiol. Cell. Physiol. 289:C293–301.CrossRefGoogle Scholar
  7. 7.
    Lentschat A et al. (2005) Mastoparan, a G protein agonist peptide, differentially modulates TLR4- and TLR2-mediated signaling in human endothelial cells and murine macrophages. J. Immunol. 174:4252–61.CrossRefGoogle Scholar
  8. 8.
    Triantafilou K, Triantafilou M, Dedrick RL. (2001) A CD14-independent LPS receptor cluster. Nat. Immunol. 2:338–45.CrossRefGoogle Scholar
  9. 9.
    Xiao YQ et al. (2006) Oxidants selectively reverse TGF-beta suppression of proinflammatory mediator production. J. Immunol. 176:1209–17.CrossRefGoogle Scholar
  10. 10.
    Xiao YQ et al. (2002) Cross-talk between ERK and p38 MAPK mediates selective suppression of pro-inflammatory cytokines by transforming growth factor-beta. J. Biol. Chem. 277:14884–93.CrossRefGoogle Scholar
  11. 11.
    Chi H et al. (2006) Dynamic regulation of pro- and anti-inflammatory cytokines by MAPK phosphatase 1 (MKP-1) in innate immune responses. Proc. Natl. Acad. Sci. U. S. A. 103:2274–9.CrossRefGoogle Scholar
  12. 12.
    Zhao Q et al. (2006) MAP kinase phosphatase 1 controls innate immune responses and suppresses endotoxic shock. J. Exp. Med. 203:131–40.CrossRefGoogle Scholar
  13. 13.
    Barber SA, Perera PY, McNally R, Vogel SN. (1995) The serine/threonine phosphatase inhibitor, calyculin A, inhibits and dissociates macrophage responses to lipopolysaccharide. J. Immunol. 155:1404–10.PubMedGoogle Scholar
  14. 14.
    Fernando LP, Fernando AN, Ferlito M, Halushka PV, Cook JA. (2000) Suppression of Cox-2 and TNF-alpha mRNA in endotoxin tolerance: effect of cycloheximide, antinomycin D, and okadaic acid. Shock. 14:128–33.CrossRefGoogle Scholar
  15. 15.
    Zhang P, Nelson S, Summer WR, Spitzer JA. (2000) Serine/threonine phosphorylation in cellular signaling for alveolar macrophage phagocytic response to endotoxin. Shock. 13:34–40.CrossRefGoogle Scholar
  16. 16.
    Francois S et al. (2005) Inhibition of neutrophil apoptosis by TLR agonists in whole blood: involvement of the phosphoinositide 3-kinase/Akt and NF-kappaB signaling pathways, leading to increased levels of Mcl-1, A1, and phosphorylated Bad. J. Immunol. 174:3633–42.CrossRefGoogle Scholar
  17. 17.
    Guha M, Mackman N. (2002) The phosphatidylinositol 3-kinase-Akt pathway limits lipopolysaccharide activation of signaling pathways and expression of inflammatory mediators in human monocytic cells. J. Biol. Chem. 277:32124–32.CrossRefGoogle Scholar
  18. 18.
    Eichholtz T, Jalink K, Fahrenfort I, Moolenaar WH. (1993) The bioactive phospholipid lysophosphatidic acid is released from activated platelets. Biochem. J. 291:677–80.CrossRefGoogle Scholar
  19. 19.
    Balazs L, Okolicany J, Ferrebee M, Tolley B, Tigyi G. (2001) Topical application of the phospholipid growth factor lysophosphatidic acid promotes wound healing in vivo. Am. J. Physiol. Regul. Integr. Comp. Physiol. 280:R466–72.CrossRefGoogle Scholar
  20. 20.
    Graler MH, Goetzl EJ. (2002) Lysophospholipids and their G protein-coupled receptors in inflammation and immunity. Biochim. Biophys. Acta 1582:168–74.CrossRefGoogle Scholar
  21. 21.
    Gueguen G et al. (1999) Structure-activity analysis of the effects of lysophosphatidic acid on platelet aggregation. Biochemistry. 38:8440–50.CrossRefGoogle Scholar
  22. 22.
    Contos JJ, Ishii I, Chun J. (2000) Lysophosphatidic acid receptors. Mol. Pharmacol. 58:1188–96.CrossRefGoogle Scholar
  23. 23.
    Moolenaar WH, van Meeteren LA, Giepmans BN. (2004) The ins and outs of lysophosphatidic acid signaling. Bioessays. 26:870–81.CrossRefGoogle Scholar
  24. 24.
    Noguchi K, Ishii S, Shimizu T. (2003) Identification of p2y9/GPR23 as a novel G protein-coupled receptor for lysophosphatidic acid, structurally distant from the Edg family. J. Biol. Chem. 278:25600–6.CrossRefGoogle Scholar
  25. 25.
    McIntyre TM et al. (2003) Identification of an intracellular receptor for lysophosphatidic acid (LPA): LPA is a transcellular PPARgamma agonist. Proc. Natl. Acad. Sci. U. S. A. 100:131–6.CrossRefGoogle Scholar
  26. 26.
    Murch O, Collin M, Thiemermann C. (2007) Lysophosphatidic acid reduces the organ injury caused by endotoxemia-a role for G-protein-coupled receptors and peroxisome proliferator-activated receptor-gamma. Shock. 27:48–54.CrossRefGoogle Scholar
  27. 27.
    Zingarelli B et al. (2002) Absence of inducible nitric oxide synthase modulates early reperfusion-induced NF-kappaB and AP-1 activation and enhances myocardial damage. Faseb. J. 16:327–42.CrossRefGoogle Scholar
  28. 28.
    Kaplan JM et al. (2005) 15-Deoxy-delta(12,14)-prostaglandin J(2) (15D-PGJ(2)), a peroxisome proliferator activated receptor gamma ligand, reduces tissue leukosequestration and mortality in endotoxic shock. Shock. 24:59–65.CrossRefGoogle Scholar
  29. 29.
    Shahrestanifar M, Fan X, Manning DR. (1999) Lysophosphatidic acid activates NF-kappaB in fibroblasts. A requirement for multiple inputs. J. Biol. Chem. 274:3828–33.CrossRefGoogle Scholar
  30. 30.
    Zingarelli B, Cook JA. (2005) Peroxisome proliferator-activated receptor-gamma is a new therapeutic target in sepsis and inflammation. Shock. 23:393–9.CrossRefGoogle Scholar
  31. 31.
    Gesty-Palmer D, El Shewy H, Kohout TA, Luttrell LM. (2005) beta-Arrestin 2 expression determines the transcriptional response to lysophosphatidic acid stimulation in murine embryo fibroblasts. J. Biol. Chem. 280:32157–67.CrossRefGoogle Scholar
  32. 32.
    Luttrell LM, Daaka Y, Lefkowitz RJ. (1999) Regulation of tyrosine kinase cascades by G-protein-coupled receptors. Curr. Opin. Cell. Biol. 11:177–83.CrossRefGoogle Scholar
  33. 33.
    Yang H, Young DW, Gusovsky F, Chow JC. (2000) Cellular events mediated by lipopolysaccharide-stimulated toll-like receptor 4. MD-2 is required for activation of mitogen-activated protein kinases and Elk-1. J. Biol. Chem. 275:20861–6.CrossRefGoogle Scholar
  34. 34.
    Koide N et al. (2005) Inhibition of extracellular signal-regulated kinase 1/2 augments nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophage cells. FEMS Immunol. Med. Microbiol. 45:213–9.CrossRefGoogle Scholar
  35. 35.
    Ruiz PA, Kim SC, Sartor RB, Haller D. (2004) 15-deoxy-delta12,14-prostaglandin J2-mediated ERK signaling inhibits gram-negative bacteria-induced RelA phosphorylation and interleukin-6 gene expression in intestinal epithelial cells through modulation of protein phosphatase 2A activity. J. Biol. Chem. 279:36103–11.CrossRefGoogle Scholar
  36. 36.
    Aga M et al. (2004) Evidence for nucleotide receptor modulation of cross talk between MAP kinase and NF-kappa B signaling pathways in murine RAW 264.7 macrophages. Am. J. Physiol. Cell. Physiol. 286:C923–30.CrossRefGoogle Scholar
  37. 37.
    Watters JJ et al. (2002) A differential role for the mitogen-activated protein kinases in lipopolysaccharide signaling: the MEK/ERK pathway is not essential for nitric oxide and interleukin 1beta production. J. Biol. Chem. 277:9077–87.CrossRefGoogle Scholar
  38. 38.
    Mayer AM, Brenic S, Stocker R, Glaser KB. (1995) Modulation of superoxide generation in in vivo lipopolysaccharide-primed rat alveolar macrophages by arachidonic acid and inhibitors of protein kinase C, phospholipase A2, protein serine-threonine phosphatase(s), protein tyrosine kinase(s) and phosphatase(s). J. Pharmacol. Exp. Ther. 274:427–36.PubMedGoogle Scholar
  39. 39.
    Sung SJ, Walters JA. (1993) Stimulation of interleukin-1 alpha and interleukin-1 beta production in human monocytes by protein phosphatase 1 and 2A inhibitors. J. Biol. Chem. 268:5802–9.PubMedGoogle Scholar
  40. 40.
    Fukao T, Koyasu S. (2003) PI3K and negative regulation of TLR signaling. Trends. Immunol. 24:358–63.CrossRefGoogle Scholar
  41. 41.
    Peck OM et al. (2006) The phosphatidylinositol 3 kinase pathway regulates tolerance to lipopolysaccharide and priming responses to Staphylococcus aureus and lipopolysaccharide. Shock. 26:31–6.CrossRefGoogle Scholar
  42. 42.
    Chou CH et al. (2005) Up-regulation of interleukin-6 in human ovarian cancer cell via a Gi/PI3K-Akt/NF-kappaB pathway by lysophosphatidic acid, an ovarian cancer-activating factor. Carcinogenesis. 26:45–52.CrossRefGoogle Scholar
  43. 43.
    Deng W et al. (2007) The lysophosphatidic acid type 2 receptor is required for protection against radiation-induced intestinal injury. Gastroenterology. 132:1834–51.CrossRefGoogle Scholar
  44. 44.
    Fromm C, Coso OA, Montaner S, Xu N, Gutkind JS. (1997) The small GTP-binding protein Rho links G protein-coupled receptors and G alpha12 to the serum response element and to cellular transformation. Proc. Natl. Acad. Sci. U. S. A. 94:10098–103.CrossRefGoogle Scholar
  45. 45.
    Sautin YY, Crawford JM, Svetlov SI. (2001) Enhancement of survival by LPA via Erk1/Erk2 and PI 3-kinase/Akt pathways in a murine hepatocyte cell line. Am. J. Physiol. Cell. Physiol. 281:C2010–9.CrossRefGoogle Scholar
  46. 46.
    Bommakanti RK, Vinayak S, Simonds WF. (2000) Dual regulation of Akt/protein kinase B by heterotrimeric G protein subunits. J. Biol. Chem. 275:38870–6.CrossRefGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2008

Authors and Affiliations

  • Hongkuan Fan
    • 1
  • Basilia Zingarelli
    • 2
  • Vashaunta Harris
    • 1
  • George E. Tempel
    • 1
  • Perry V. Halushka
    • 3
  • James A. Cook
    • 1
  1. 1.Department of NeurosciencesMedical University of South CarolinaCharlestonUSA
  2. 2.Division of Critical Care MedicineCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  3. 3.Medicine, and PharmacologyMedical University of South CarolinaCharlestonUSA

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