Analysis of inflammatory and lipid metabolic networks across RAW264.7 and thioglycolate-elicited macrophages

Mano R. Maurya, Shakti Gupta, Xiang Li, Eoin Fahy, Ashok R. Dinasarapu, Manish Sud, H. Alex Brown, Christopher K. Glass, Robert C. Murphy, David W. Russell, Edward A. Dennis, Shankar Subramaniam

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Studies of macrophage biology have been signifi- cantly advanced by the availability of cell lines such as RAW264.7 cells. However, it is unclear how these cell lines differ from primary macrophages such as thioglycolateelicited peritoneal macrophages (TGEMs). We used the inflammatory stimulus Kdo 2 -lipid A (KLA) to stimulate RAW264.7 and TGEM cells. Temporal changes of lipid and gene expression levels were concomitantly measured and a systems-level analysis was performed on the fold-change data. Here we present a comprehensive comparison between the two cell types. Upon KLA treatment, both RAW264.7 and TGEM cells show a strong inflammatory response. TGEM (primary) cells show a more rapid and intense inflammatory response relative to RAW264.7 cells. DNA levels (fold-change relative to control) are reduced in RAW264.7 cells, correlating with greater downregulation of cell cycle genes. The transcriptional response suggests that the cholesterol de novo synthesis increases considerably in RAW264.7 cells, but 25-hydroxycholesterol increases considerably in TGEM cells. Overall, while RAW264.7 cells behave similarly to TGEM cells in some ways and can be used as a good model for inflammation- and immune function- related kinetic studies, they behave differently than TGEM cells in other aspects of lipid metabolism and phenotypes used as models for various disorders such as atherosclerosis.

Original languageEnglish (US)
Pages (from-to)2525-2542
Number of pages18
JournalJournal of lipid research
Volume54
Issue number9
DOIs
StatePublished - Sep 2013

Keywords

  • Cell cycle
  • Correlation analysis
  • Inflammation
  • Lipid and signaling networks
  • Lipidomics
  • Primary macrophage
  • Toll-like receptor 4
  • Transcriptomics

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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  • Cite this

    Maurya, M. R., Gupta, S., Li, X., Fahy, E., Dinasarapu, A. R., Sud, M., Brown, H. A., Glass, C. K., Murphy, R. C., Russell, D. W., Dennis, E. A., & Subramaniam, S. (2013). Analysis of inflammatory and lipid metabolic networks across RAW264.7 and thioglycolate-elicited macrophages. Journal of lipid research, 54(9), 2525-2542. https://doi.org/10.1194/jlr.M040212