Correlation between endotoxin tolerance in human monocyte leukemia cell line THP-1 with glucocorticoid receptor-α

Abstract

Human monocyte leukemia cell line THP-1 was stimulated with lipopolysaccharide (LPS) to simulate the sepsis model and the expression of human glucocorticoid receptor-α (GR-α) mRNA in montocytes with endotoxin tolerance was investigated. THP-1 cells were cultured in serum-free medium, randomly divided into groups A, B, C, D and E, and stimulated with 0, 10, 10, 100,0 ng/mL LPS for 24 h followed with 100, 100, 10, 100,0 ng/mL LPS for another 24 h respectively. The expression of GR-α mRNA was detected by semi-quantitative reverse transcriptional polymerase chain reaction. Tumor necrosis factor-α (TNF-α) was determined by enzyme linked immunosorbent assay (ELISA). The results showed that the A values of GR-α/β-actin in groups A, B, C, D and E was, 0.607±0.006, 0.368±0.005, 0.484±0.008, 0.509±0.004 and 0.564±0.014 respectively with the difference being significant among the groups (P<0.05). The GR-α mRNA expression was negatively correlated with the TNF-α expression (P|<0.01). It was concluded that the down-regulation of the expression of GR-α mRNA in endotoxin tolerance THP-1 cells might play an important role in the development of endotoxin tolerance in THP-1 cells.

Keywords

endotoxin tolerance / sepsis / THP-1 cell / glucocorticoid receptor

References

[1] NomuraF, AkashiS, SakaoY, et al.. Cutting edge: Endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface toll-like receptor 4 expression. J Immunol, 2000, 164: 3476-3479

[2] HollenbergS M, WeinbergerC, OngE S, et al.. Primary structure and expression of a functional human glucocorticoid receptor cDNA. Nature, 1985, 318(6047): 635-641

[3] ShengZ Y, YaoY M. Sepsis and multiple organ dysfunction syndrome. Chin J Emerg Med (Chinese), 2003, 12(10): 653-653

[4] BledsoeR K, MontanaV G, StanleyT B. Crystal structure of the glucocorticoid receptor ligand binding domain reveals a novel mode of receptor dimerization and coactivator recognition. Cell, 2002, 12(110): 93-105

[5] BambergerC M, BambergerA, de CastroM, et al.. Glucocorticoid receptor beta, a potential endogenous inhibitor of glucocorticoid action in humans. J Clin Invest, 1995, 95: 2435-2441

[6] OakleyR H, SarM, CidlowskiJ A. The human glucocorticoid receptor beta isoform, expression, biochemical properties, and putative function. J Biol Chem, 1996, 271: 9550-9559

[7] de CastroM, ElliotS T, BambergerC M, et al.. The non-ligand binding beta-isoform of the human glucocorticoid receptor (hGR beta): tissue levels, mechanism of action, and potential physiologic role. Mol Med, 1996, 2: 597-607

[8] Ziegler-HeitbrockH W L, WedelA, SchrautW, et al.. Tolerance to lipopolysaccharide involves mobilization of nuclear factor κB with predominance of p50 homodimers. J Biol Chem, 1994, 269: 17001-17004

[9] GoldringC E, ReveneauS, PinardD, et al.. Hypore-sponsiveness to lipopolysaccharide alters the composition of NF-kappa B binding to the regulatory regions of inducible nitric oxide synthase gene. Eur J Immunol, 1998, 28: 2960-2970

[10] KastenbauerS, Ziegler-HeitbrockH W. NF- kappa B1 (p50) is up-regulated in lipopolysaccharide tolerance and can block tumor necrosis factor gene expression. Infect Immun, 1999, 67(4): 1553-1559

[11] BarnesP J, KarinM. Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med, 1997, 336: 1066-1071