Bradykinin increases permeability by calcium and 5-lipoxygenase in the ECV304/C6 cell culture model of the blood–brain barrier

摘要: Abstract The blood–brain barrier (BBB) was modelled in this study using ECV304 cells co-culture with rat C6 glioma cells, which resulted elevated transendothelial electrical resistance (TEER). inflammatory mediator bradykinin (1 μM) studied and found to induce a fall TEER; the link between change intracellular free calcium concentration ([Ca2+]i) then examined. 1 μM produced peak–plateau increase [Ca2+]i. peak showed desensitization dose dependent (over 0.1 nM μM). [Ca2+]i blocked by B2 antagonist HOE 140 without effect from B1 agonist antagonist. plateau response abolished Ca2+-free solution containing 2 mM EDTA, also Ca2+ channel blockers lanthanum, La3+ (10 μM), SKF 96365 (100 store Ca2+ATPase inhibitor thapsigargin response. putative phospholipase C inhibitors, U73122 (20 ETH-18-OCH3 unexpectedly increased [Ca2+]i; after their application, ineffective. Agents on responses included A2 (PLA2) aristolochic acid (0.5 mM), cyclooxygenase indomethacin 5-lipoxygenase nordihydroguaiaretic acid, NDGA calphostin l -NAME mM) nifedipine TEER 140, combined La3+, NDGA, but not indomethacin, or -NAME. while reduced it. Thus through receptor-linked release of thapsigargin-sensitive stores, leading activation PLA2 metabolism arachidonic 5-lipoxygenase.