International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Inhibition of Lipopolysaccharide-stimulated Inflammatory Cytokine Production by LY303511 in Human Macrophagic THP-1 Cells

  • Kim, So-Hee (Department of Oral Microbiology, School of Dentistry, Chonnam National University) ;
  • Paek, Yun-Woong (Department of Physical Therapy, Gwangju Health University) ;
  • Kang, In-Chol (Department of Oral Microbiology, School of Dentistry, Chonnam National University)
  • Received : 2017.08.18
  • Accepted : 2017.09.05
  • Published : 2017.09.30
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Abstract

We have previously shown that the specific phosphatidylinositol 3-kinase inhibitor LY294002 (LY29), and its inactive analog LY303511 (LY30), inhibit a monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells; these results suggest the potential of LY30 as an anti-inflammatory drug. In this study, we determined the effects of LY30 on the production of various inflammatory cytokines in human macrophagic THP-1 cells which were stimulated with lipopolysaccharide (LPS). LY30 selectively suppressed the mRNA expression of IL-12 p40, $TNF-{\alpha}$, and MCP-1 without affecting the expression of $IL-1{\alpha}$, IL-6, and IL-8. Inhibition of the production of IL-12 and $TNF-{\alpha}$ by LY30 was also demonstrated using ELISA assays. In order to elucidate the mechanisms of the action of LY30, we examined the role played by the mitogen-activated protein kinases and the key transcription factors, AP-1 and $NF-{\kappa}B$ in LPS-stimulated THP-1 cells. The results revealed that LY30 inhibited LPS-induced activation of ERK, but not p38 or JNK. Furthermore, the AP-1 DNA binding activity was suppressed by LY30 based upon the dosage, whereas $NF-{\kappa}B$ DNA binding was not affected. These results suggest that LY30 selectively inhibits cytokine production in the LPS-stimulated macrophagic THP-1 cells by down-regulating the activation of ERK and AP-1.

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References

  1. Adi S, Wu NY, Rosenthal SM. Growth factor-stimulated phosphorylation of Akt and p70(S6K) is differentially inhibited by LY294002 and Wortmannin. Endocrinology. 2001;142:498-501. doi: 10.1210/endo.142.1.8051.
  2. Arcaro A, Wymann MP. Wortmannin is a potent phosphatidylinositol 3-kinase inhibitor: the role of phosphatidylinositol 3,4,5-trisphosphate in neutrophil responses. Biochem J. 1993;296:297-301. https://doi.org/10.1042/bj2960297
  3. Vlahos CJ, Matter WF, Hui KY, Brown RF. A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem. 1994;269:5241-5248.
  4. Choi EK, Park HJ, Ma JS, Lee HC, Kang HC, Kim BG, Kang IC. LY294002 inhibits monocyte chemoattractant protein-1 expression through a phosphatidylinositol 3-kinaseindependent mechanism. FEBS Lett. 2004;559:141-144. doi: 10.1016/s0014-5793(04)00058-4.
  5. Kristof AS, Pacheco-Rodriguez G, Schremmer B, Moss J. LY303511 (2-piperazinyl-8-phenyl-4H-1-benzopyran-4-one) acts via phosphatidylinositol 3-kinase-independent pathways to inhibit cell proliferation via mammalian target of rapamycin (mTOR)- and non-mTOR-dependent mechanisms. J Pharmacol Exp Ther. 2005;314:1134-1143. doi: 10.1124/jpet.105.083550.
  6. Poh TW, Pervaiz S. LY294002 and LY303511 sensitize tumor cells to drug-induced apoptosis via intracellular hydrogen peroxide production independent of the phosphoinositide 3-kinase-Akt pathway. Cancer Res. 2005;65:6264-6274. doi: 10.1158/0008-5472.can-05-0152
  7. Avni D, Glucksam Y, Zor T. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 modulates cytokine expression in macrophages via p50 nuclear factor-${\kappa}B$ inhibition, in a PI3K-independent mechanism. Biochem Pharmacol. 2012;83:106-114. doi: 10.1016/j.bcp.2011.09.025.
  8. Tas SW, Baeten DL. Recent Advances in the Treatment of Immune-Mediated Inflammatory Diseases. Methods Mol Biol. 2016;1371:143-155. doi: 10.1007/978-1-4939-3139-2_9.
  9. Arango Duque G, Descoteaux A. Macrophage cytokines: involvement in immunity and infectious diseases. Front Immunol. 2014;5:491. doi: 10.3389/fimmu.2014.00491.
  10. Auwerx J. The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiation. Experientia. 1991;47:22-31. https://doi.org/10.1007/BF02041244
  11. Zhang YL, Dong C. MAP kinases in immune responses. Cell Mol Immunol. 2005;2:20-27.
  12. Adcock IM. Transcription factors as activators of gene transcription: AP-1 and $NF-{\kappa}B$. Monaldi Arch Chest Dis. 1997;52:178-186.
  13. Vallabhapurapu S, Karin M. Regulation and function of $NF-{\kappa}B$ transcription factors in the immune system. Annu Rev Immunol. 2009;27:693-733. doi: 10.1146/annurev.immunol.021908.132641.
  14. Choi EK, Jang HC, Kim JH, Kim HJ, Kang HC, Paek YW, Lee HC, Lee SH, Oh WM, Kang IC. Enhancement of cytokinemediated $NF-{\kappa}B$ activation by phosphatidylinositol 3-kinase inhibitors in monocytic cells. Int Immunopharmacol. 2006;6:908-915. doi: 10.1016/j.intimp.2006.01.007.
  15. Roskoski R, Jr. ERK1/2 MAP kinases: structure, function, and regulation. Pharmacol Res. 2012;66:105-143. doi: 10.1016/j.phrs.2012.04.005.