NBD-cholesterol incorporation by rat macrophages and lymphocytes: a process dependent on the activation state of the cells

Cell Biochem Funct. 2004 Jan-Feb;22(1):23-8. doi: 10.1002/cbf.1048.

Abstract

The time-course of incorporation of NBD-cholesterol by macrophages (Ma) and lymphocytes (LY) obtained from untreated and thioglycollate-injected (thio) rats was investigated. NBD-cholesterol incorporation was also examined in Ma obtained from untreated rats and stimulated in vitro by lipopolysaccharide (LPS) and phorbol-myristate acetate (PMA). The same measurement was performed in LY from untreated rats stimulated by addition of LPS and concanavalin A (Con A) into the culture medium. Thio-treated Ma showed high fluorescence intensity after 1 h of incubation with NBD-cholesterol. Ma submitted concomitant to LPS and NBD-cholesterol showed low fluorescence intensity, as well as Ma stimulated with PMA. Ma from untreated and LPS pre-treated rats showed a similar time-course of incorporation. LY from thio-treated rats showed lower incorporation of NBD-cholesterol in comparison to LY from untreated rats. Incorporation was reduced when LPS was added concomitantly with NBD-cholesterol. On the other hand, LY pre-treated with LPS for 48 h showed a very high incorporation of NBD-cholesterol. Con A treatment did not cause a significant effect on NBD-cholesterol incorporation. The findings presented herein led us to conclude that the uptake of NBD-cholesterol by Ma and LY is markedly affected by the activation state of the cells.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • 4-Chloro-7-nitrobenzofurazan / metabolism*
  • Animals
  • Cholesterol / analogs & derivatives*
  • Cholesterol / metabolism*
  • Concanavalin A / pharmacology
  • Lipopolysaccharides / pharmacology
  • Lymphocyte Activation / drug effects
  • Lymphocytes / drug effects
  • Lymphocytes / metabolism*
  • Macrophage Activation / drug effects
  • Macrophages, Peritoneal / drug effects
  • Macrophages, Peritoneal / metabolism*
  • Male
  • Rats
  • Rats, Wistar
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • 7-nitrobenz-2-oxa-1,3-diazol-4-ylcholesterol
  • Lipopolysaccharides
  • Concanavalin A
  • Cholesterol
  • 4-Chloro-7-nitrobenzofurazan
  • Tetradecanoylphorbol Acetate