Characteristics of histamine-induced leukocyte rolling in the undisturbed microcirculation of the rat mesentery

1 The main objective of this study was to analyse the role and mode of action of the mast cell mediator histamine in leukocyte-endothelium interactions in small venules in vivo. For this put-pose. we used a histological approach (combined with intravital microscopy) that allows studies of rapid mediator-induced venular leukocyte accumulation, reflecting leukocyte rolling, in the undisturbed microcirculation of the rat mesentery where rolling is normally absent. 2 We first examined the relative importance of histamine and 5-hydroxytryptamine (5-HT) in acute mast cell-dependent leukocyte recruitment. The mast cell secretagogue compound 48/80 (i.p. for 15 min) induced a marked venular accumulation of polymorphonuclear leukocytes (PMNL) which was almost abolished by combined histamine(1) (H-1)- and histamine(2) (H-2)-receptor blockade. In contrast, the 5-HT- receptor antagonist methysergide was inactive in this regard. Moreover, exogenous 5-HT was less active than exogenous histamine in evoking venular PMNL accumulation (histamine response dose-dependent; 5-HT response bell shaped). Prostaglandin D-2 did not cause PMNL accumulation. 3 The venular PMNL response to exogenous histamine peaked between 15 min and 1 h. was still significantly elevated at 2 hi and then returned to prechallenge values after 3 h. At all time points, the histamine-induced PMNL accumulation was nearly abolished by i.v, treatment with the polysaccharide fucoidin (which blocks rolling but not firm adhesion per se), suggesting that the PMNL response to histamine was due to rolling rather than firm adhesion over the entire 3 h period. At no time Feint did histamine trigger accumulation of mononuclear leukocytes (MNL). 4 To examine the role of histamine-receptors in the histamine-induced PMNL accumulation (i.e. rolling). the animals were pretreated with diphenhydramine (Hi-receptor antagonist), cimetidine. or ranitidine (H-2-receptor antagonists). Diphenhydramine alone inhibited the venular PMNL response to histamine by 52%, while both H-2-receptor antagonists were completely inactive. However, the combination of cimetidine and diphenhydramine reduced the histamine-induced PMNL rolling by 82%. Furthermore? in contrast to an H-3-receptor agonist challenge with either the H-1-receptor agonist 2-thiazolylethylamine or two different H-2-receptor agonists (impromidine, dimaprit) was sufficient to provoke significant venular PMNL accumulation. 5 Treatment with the nitric oxide-synthase inhibitor L-NAME did not affect the histamine-induced PMNL rolling. On the other hand, 3 h pretreatment with dexamethasone reduced the PMNL response to histamine by 73%, and flow cytometric analysis showed that the dexamethasone treatment almost completely inhibited binding of soluble P-selectin to rat isolated PMNLs. 6 We conclude that initial leukocyte recruitment after mast cell activation in the rat mesentery is critically dependent on histamine release. The cellular response to histamine was specifically due to PMNL rolling, involved activation of both H-1- and H-2-receptors, and lasted for 2-3 h. Moreover? the histamine-induced PMNL rolling was not dependent on nitric oxide synthesis, but was sensitive to glucocorticoid treatment, possibly via inhibition of expression or function of leukocytic P-selectin ligand(s).