期刊
CELL CALCIUM
卷 72, 期 -, 页码 116-126出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.ceca.2018.02.002
关键词
Actin; BAPTA; Cell barrier; Fura-2; Histamine; Inflammation; Intracellular calcium; Mast cell; Microtubules; Peritoneal dialysis; Permeability; Serosal membrane
类别
资金
- Baxter Healthcare, USA
- Fresenius Medical Care, Germany, GmbH
Mast cells in the peritoneal membrane (PM) may degranulate to release preformed inflammatory mediators including histamine which is capable of diffusing into the surrounding interstitium, modulating cells in their vicinity including, human peritoneal mesothelial cells (hPMC). The present study aimed to investigate the quorum intracellular calcium ([Ca-i(2+)]) response to histamine compared to the membrane soluble ionophore, A23187, in adherent cultured hPMC. To examine [Ca-i(2+)] handling, Fura - 2 loaded cells were exposed to histamine and A23187. Agonist induced transient [Ca-i(2+)] event(s) (TCE) were defined and compared including, resting calcium, peak height, recovery and transient kinetics. Changes in cell shape were examined with immunocytochemistry of the cortical actin (CA) and microtubule (MT) cytoskeleton. To investigate whether histamine induced changes in cell shape were mediated by [Ca-i(2+)], mobilization of [Ca-i(2+)] was prevented with 20[mu mol/l of the calcium chelator l,2-bis-(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Histamine produced a dose dependent increase of [Ca-i(2+)], maximal at 1.0 mmol/1 which recovered to the pre-challenge resting value. Transient multiplicity with repeated challenge was evident below a histamine threshold of 10(-2) mmol/1. Morphometric analysis of MTs and CA showed significant cell elongation plus histamine and A23187. The histamine induced cell elongation was eliminated with [Ca-i(2+)] clamping. This data indicated that increased [Ca-i(2+)] was essential for cell elongation and the formation of stress fibres and therefore has a pivotal role in the regulation of the PM barrier.
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