title: Effect of dehydroepiandrosterone on lipopolysaccharide-induced interleukin-6 production in DH82 cultured canine macrophage cells.
abstract: OBJECTIVE
  This study examined the effect of dehydroepiandrosterone (DHEA) on lipopolysaccharide (LPS)-stimulated interleukin-6 (IL-6) production in the DH82 canine macrophage cell line.
  
  
  METHODS
  Cultured DH82 cells were stimulated with varying concentrations of LPS with or without DHEA for various times. Supernatant IL-6 levels were measured by enzyme-linked immunosorbent assay, and cellular cytoplasmic IkappaBalpha protein expression measured by Western blot analysis.
  
  
  RESULTS
  LPS dose-dependently stimulated IL-6 production (p=0.016). Cells stimulated with 20 microg LPS showed a time-dependent increase of IL-6 concentration up to 10 h post-treatment (p=0.007). Co-treatment of DH82 cells with 20 microg LPS and various concentrations of DHEA for 14 h showed that up to 10 microM DHEA dose-dependently decreased the IL-6 concentration (p=0.007). Also, addition of 20 microM DHEA to DH82 cells with 20 microg LPS time-dependently decreased the IL-6 concentration for up to 14 h post-treatment (p=0.018). Stimulation of cultured DH82 cells with 20 microg LPS significantly decreased cellular cytoplasmic IkappaBalpha expression, beginning at 30 min post-treatment and persisting to at least 2 h post-treatment (p=0.012). However, co-treatment of cells with 20 microg LPS and 20 microM DHEA abrogated this effect until 2 h post-treatment.
  
  
  CONCLUSIONS
  DHEA decreased the IL-6 concentration in the supernatant of LPS-stimulated DH82 cells by inhibiting the sequestration of IkappaBalpha, which is necessary for the activation of nuclear factor-kappa B. These findings provide new insights into the immunomodulatory effects of DHEA.
authors: Department of Obstetrics and Gynecology, University of Ulsan College of Medicine, Asan Medical Center, 388-1 Pungnap-dong Songpa-gu, Seoul 138-736, Republic of Korea.
authors: Sun-Kwon
authors: SK
authors: Kim
authors: Myung-Shin
authors: MS
authors: Shin
authors: Bok-Kyung
authors: BK
authors: Jung
authors: Jae-Yoon
authors: JY
authors: Shim
authors: Hye-Sung
authors: HS
authors: Won
authors: Pil Ryang
authors: PR
authors: Lee
authors: Ahm
authors: A
authors: Kim