Temporal regulation by adrenergic receptor stimulation of macrophage (M phi)-derived tumor necrosis factor (TNF) production post-LPS challenge

Macrophage (M phi) responsiveness can be regulated by various mediators, including those which emanate from, and mimic, the sympathetic nervous system. Whereas beta-adrenergic agonists suppress, alpha(2)-adrenergic agonists augment lipopolysaccharide (LPS)-stimulated tumor necrosis factor (TNF) production and gene expression. The susceptibility of M phi s to regulation of LPS-induced TNF production and mRNA accumulation was examined following beta-adrenergic and alpha(2)-adrenergic receptor activation at specific time points post-LPS challenge. Complete Freund's adjuvant-elicited murine M phi s were incubated with LPS (30 ng/ml) in the presence or absence of adrenergic agonists or antagonists. We assessed the susceptibility of immunologically-activated M phi s to adrenergic receptor regulation: a) during the 1 h delay in the production of TNF after LPS-stimulation, and b) during the rapid increase in TNF production which follows. Disparate responsiveness of M phi s to adrenergic drugs was observed during this time course of TNF production and TNF mRNA accumulation. In particular, while the concomitant addition of an az-adrenergic antagonist and LPS resulted in 45% suppression of TNF production, this selective blockade of alpha(2)-adrenergic receptors on M phi s was equally effective throughout the first 45 min post-LPS challenge. After this initial period, the alpha(2)-adrenergic receptor became progressively less responsive as demonstrated by the delayed addition of yohimbine (10(-5) M) post-LPS challenge. The addition of the selective alpha(2)-adrenergic agonist UK-14304 (10(-7) M) to LPS-activated M phi s augmented TNF mRNA accumulation. However, this augmentation was even greater when the addition of the alpha(2)-adrenergic agonist was delayed post-LPS challenge. It was also shown that the beta-adrenergic agonist isoproterenol (10(-6) M) produced maximum suppression of TNF production within the first 1.5 h post-LPS challenge. Suppression by isoproterenol (10(-6) M) of TNF mRNA accumulation occurred throughout the 2-h period assessed post-LPS stimulation of M phi s. The decline in isoproterenol-induced regulation was accompanied by an elevation in beta(2)-adrenergic receptor mRNA accumulation. Furthermore, suppression of TNF production induced by a maximum concentration of isoproterenol was observed at various LPS concentrations (0.001-1000 ng/ml), although this was not as pronounced a suppression as demonstrated for dibutyryl cAMP. These results demonstrate that the susceptibility of M phi s to adrenergic receptor regulation changes throughout the time period necessary for gene activation and ultimate release of TNF. Thus, the production of TNF during LPS-dependent disease states may be regulated by adrenergic mediators throughout different temporal windows, better explaining the role played by the nervous system.