DOI QR코드

DOI QR Code

Proinflammatory Effects of Bacterial Lipopolysaccharide (LPS) in Rainbow Trout (Oncorhynchus mykiss) Macrophage Cells

  • Hong Suhee (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Jeong Hyun Do (Department of Aquatic Life Medicine, Pukyong National University)
  • Published : 2003.09.01

Abstract

Proinflammatory effects of bacterial lipopolysaccharide (LPS) have been assessed by analysing the induction of two inflammatory genes, $interleukin-1\beta$ $(IL-1\beta)$ and cyclooxygenase-2 (COX-2), in rainbow trout (Oncorhynchus mykiss) macrophage cells. Production of a metabolite of arachidonic acid by COX-2, prostaglandin $E_2\;(PGE_2)$, was also analysed in macrophage cells after LPS stimulation. Northern blot analysis revealed that LPS $(5{\mu}g/mL)$ significantly upregulated $IL-1\beta$ (54 times) and COX-2 (40.7 times) gene expression in macrophage cells after 4 h stimulation. According to RT-PCR (Reverse Transcription Polymerase Chain Reaction) analysis, $IL-1\beta$ gene induction in LPS stimulated macrophage cells was started within 1h and significantly increased thereafter until 4h. Meanwhile, COX-2 gene induction by LPS was delayed in comparison with $IL-1\beta$ gene induction as a faint band was observed after 4h stimulation in head kidney macrophage cells. LPS also significantly increased $PGE_2$ production in head kidney leucocytes, presumably via activating COX-2 expression that metabolites arachidonic acid to $PGE_2$. In conclusion, it was demonstrated that LPS could induce two main inflammatory and immune related genes, $IL-1\beta$ and COX-2, and increase $PGE_2$ production in trout head kidney macrophage cells, representing a strong inflammatory activity.

Keywords

References

  1. Auger, M.J. and J.A. Ross. 1992. The Biology of the Macrophage. In: The Macrophage, Lewis, C.E., J.O'D., McGee, eds. IRL Press, Oxford, pp. 1-74
  2. Bayne, C.J. Pahgocytosis and non-self recognition in invertebrates. 1990. Bioscience, 40, 723-731 https://doi.org/10.2307/1311504
  3. Diaz, A., K.P. Chepenik, J.H. Kom, A.M. Reginato and S.A. Jimenez. 1998. Differential regulation of cycloxygenases 1 and 2 by interluekin-I beta, tumor necrosis factor-alpha, and transforming growth factor-beta 1 in human lung fibroblasts. Exp. Cell Res., 241, 222-229 https://doi.org/10.1006/excr.1998.4050
  4. Dinarello, C.A. 1984. Interleukin I. Rev. Infect. Dis., 6, 51-95 https://doi.org/10.1093/clinids/6.1.51
  5. Fenton, M.J., M.W. Vermeulen, B.D. Clark, A.C. Webb and P.E. Auron. 1988. Human pro-IL-1beta gene expression in monocytic cells is regulated by two distinct pathways. J. lmmunol., 140, 2267-2273
  6. Fenton, M.J. 1992. Review: Transcriptional and post-transcriptional regulation of interleukin 1 gene regulation. Int. J. Immunopharmacol., 14, 401-411 https://doi.org/10.1016/0192-0561(92)90170-P
  7. Holland, J. 1997. The generation of eicosanoids in rainbow trout with particular reference to gills and leukocytes. Ph.D thesis, University of Wales, pp. 110-112
  8. Jarrous, N. and R. Kaempfer. 1994. Induction of human interleukin gene expression by retinoic acid and its regulation at processing of precursor transcripts. J. Biol. Chem., 269, 23141-23149
  9. Komaki, G., A. Arimura and K. Koves. 1992. Effects of intravenous injection of IL-1beta on $PGE_2$ levels in several brain areas as determined by microdialysis. Am. J. Physiol. 262, E246-E251
  10. Sergey, E.L., D. Gayle, M.C. Flynn and e.R. Plata-Salaman. 1998. IL-I$\beta$ system (ligand, receptor type I, receptor accessory protein and receptor antagonist), TNF $\alpha$, TGF-$\beta$ and neuropeptide Y mRNAs in specific brain regions during bacterial LPS-induced anorexia. Brain Res. Bull., 45, 507-515 https://doi.org/10.1016/S0361-9230(97)00437-1
  11. Wang, T., J Zou, C. Cunningham and C.J. Secombes. 2002. Cloning and functional characterisation of the interleukin-l$\beta$I promoter of rainbow trout. Biochim. Biophys. Acta, 1575, 108-116 https://doi.org/10.1016/S0167-4781(02)00235-X
  12. Zou, J., P.S. Grabowski, e. Cunningham and C.J. Secombes. 1998. Molecular cloning of interleukin-1beta from rainbow trout reveals no evidence of an ice cut site. Cytokine, 11, 552-560 https://doi.org/10.1006/cyto.1998.0470
  13. Zou, L, N.F. Neumann, J.W. Holland, M.S. Bellosevic, C. Cunningham, C.J. Secombes and A.F. Rowley. 1999. Fish macrophage express a cyclo-oxygenase-2 homologue after activation. Biochem. J., 340, 153-159 https://doi.org/10.1042/0264-6021:3400153