Effects of physiological concentrations of steroid hormones and interleukin-11 on basal and stimulated production of interleukin-8 by human osteoblast-like cells with different functional profiles.
OBJECTIVE
IL-8 is a CXC chemokine involved in the pathogenesis of articular damage in rheumatoid arthritis. Local hyperproduction of IL-8 has been suggested to play a role in subchondral bone loss, since it suppresses osteoblast activity and promotes osteoclasts recruitment. Osteoblasts are a source of IL-8; its secretion is regulated by a number of hormones and cytokines. The aim of the present study was to evaluate the single and combined effects of physiological concentrations of cortisol, 17 beta-estradiol and IL-11 upon basal and IL-1 beta-inducible production of IL-8 in two human osteoblast-like cell lines, Saos-2 and MG-63.


METHODS
Cells were incubated with cortisol (0.01 to 1 microM), 17 beta-estradiol (10 to 1000 pg/ml), IL-11 (1 to 100 ng/ml), in presence or absence of IL-1 beta (10 ng/ml), for 20 h. Combinations of 17 beta-estradiol and cortisol, and of IL-11 and cortisol, were also tested. After incubation, IL-8 levels in supernatants were measured by ELISA.


RESULTS
Cortisol dose-dependently inhibited spontaneous IL-8 secretion in both cell lines, although statistical significance was attained in the MG-63 cells only (P < 0.01); no effect of 17 beta-estradiol was apparent. With regard to IL-1 beta-inducible production, cortisol dose-dependently inhibited IL-8 release in both cell lines (P < 0.01); 17 beta-estradiol resulted in only a non-significant decrease in Saos-2, but not in MG-63 cells. 17 beta-estradiol did not alter the effects of cortisol in experiments involving co-incubation. IL-11 did not have any effect on spontaneous IL-8 release, but exerted a significant inhibitory effect on IL-1 beta-inducible release in MG-63 cells (P < 0.05); no additional effect was observed upon the degree of cortisol-dependent inhibition.


CONCLUSIONS
Cortisol is a potent physiological inhibitor of IL-8 production by osteoblast-like cells. The results of the present study support the use of exogenous supplemental glucocorticoids to prevent the deleterious effects of excess IL-8. The estrogenic milieu and local concentrations of IL-11 have little if any effect on the IL-8-dependent mechanisms of disease.
Dovio
A
A
Internal Medicine Unit, Department of Clinical and Biological Sciences, University of Turin, Orbassano, TO, Italy.
Sartori
M L
ML
Masera
R G
RG
Peretti
L
L
Perotti
L
L
Angeli
A
A
D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000494:pharmacology*; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology; D017370:Interleukin-11 / Q000494:pharmacology*; D016209:Interleukin-8 / Q000378:metabolism*; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D011994:Recombinant Proteins
D004338:Drug Combinations; D007375:Interleukin-1; D017370:Interleukin-11; D016209:Interleukin-8; D011994:Recombinant Proteins; D004958:Estradiol; D006854:Hydrocortisone