Mast Cell Histamine-Induced Calcium Transients in Cultured Human Peritoneal Mesothelial Cells

Abstract

ObjectivePeritoneal inflammation results from a complex interplay of events initiated by macrophage activity in response to infection, with the stimulation of mesothelial cell cytokine release amplifying the recruitment of blood-borne defense cells to the site of injury. Resident peritoneal mast cells may add to this complexity with mast cell derived cytokines released during this cascade. This study examined the influence of histamine, a mast cell derived inflammatory mediator, on the initial activation of human peritoneal mesothelial cells (HPMC) by intracellular free calcium (Cai2+) mobilization, and changes to the actin cytoskeleton.DesignHPMC signal transduction was examined in response to histamine (1.0 mmol/L) compared to fetal bovine serum (FBS) (0.1 %) and 4-br-A23187 (1.0 μmol/L). Intracellular free calcium was measured in fura-2 loaded cells with and without external calcium (Ca2ex+t), or Ca2ex+t with verapamil (100 μmol/L). Following treatment with agonists, HPMC actin cytoskeleton was stained using direct immunocytochemistry.ResultsHPMC responded to histamine with a twofold transient rise in Cai2+ which returned to the baseline, in contrast with FBS and A23187–induced Cai2+ transients, which returned to elevated resting values. In the absence of Ca2ex+t’ all agents produced a calcium transient indicative of calcium release from intracellular stores. Histamine induced calcium -dependent changes to the cytoskeleton and cellular organization, including increased actin stress fibers.ConclusionHistamine produced large specific receptor-mediated calcium transients in HPMC, which included components of calcium release from intracellular stores and receptor -mediated calcium influx processes. The observed response to histamine raises the possibility that histamine derived from resident mast cells may modulate mesothelial cell function, in part by calciumdependent pathways, and influence the performance of the peritoneal membrane during peritoneal dialysis.