Co-regulated expression of glomerular 12/15-lipoxygenase and interleukin-4 mRNAs in rat nephrotoxic nephritis

Kidney Int. 1994 Aug;46(2):341-9. doi: 10.1038/ki.1994.280.

Abstract

Arachidonate 12- and 15-lipoxygenase (LO) products are generated in experimental glomerulonephritis. 15-S-HETE (a 15-LO product) and lipoxins (interaction products between 5-LO and either 12-LO or 15-LO) counteract the proinflammatory actions of leukotrienes. IL-4 has been shown to up-regulate 15-LO gene expression in human leukocytes. Based on homology with human 15-LO, we cloned a 0.76 kbp fragment of a rat LO cDNA from leukocytes stimulated by interleukin-4 (IL-4). The deduced amino acid sequence shows 71.0% and 60.1% homology to human 15-LO and 12-LO, respectively, and 100% homology to a recently cloned "leukocyte type" rat 12-lipoxygenase enzyme, which possesses significant 15-lipoxygenase activity (heretofore referred to as "12/15-LO"). A deletion mutant was utilized to generate internal standard cRNA in quantitative PCR assays. Glomerular 12/15-LO mRNA increased significantly over controls 24 and 48 hours after NTS injection, then decreased at 72 hours. RNA from NTS glomeruli contained higher levels of 12/15-LO mRNA than that from unstimulated peripheral leukocytes, suggesting that 12/15-LO transcription is up-regulated locally in native and/or infiltrating glomerular cells. Glomerular IL-4 mRNA increased markedly 16 hours post-NTS, and was then reduced, suggesting a potential role for T cell-derived IL-4 in directing the expression of 12/15-LO during glomerulonephritis. This represents the first demonstration of tandem regulated in vivo gene expression for a lymphokine (IL-4) and a lipoxygenase, both of which promote counter-inflammatory influences in immune complex-mediated injury.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arachidonate 12-Lipoxygenase / genetics
  • Arachidonate 12-Lipoxygenase / metabolism
  • Arachidonate 15-Lipoxygenase / genetics
  • Arachidonate 15-Lipoxygenase / metabolism
  • Arachidonate Lipoxygenases / genetics
  • Arachidonate Lipoxygenases / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • Gene Expression Regulation*
  • Glomerulonephritis / enzymology*
  • Humans
  • Interleukin-4 / genetics
  • Interleukin-4 / metabolism*
  • Kidney Glomerulus / enzymology*
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Wistar
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • RNA, Messenger
  • Interleukin-4
  • Arachidonate Lipoxygenases
  • Arachidonate 12-Lipoxygenase
  • Arachidonate 15-Lipoxygenase

Associated data

  • GENBANK/S74450