Endothelins release 51Cr from cultured human cerebromicrovascular endothelium.
The effects of vasoactive peptides endothelins (ET-1, ET-2, ET-3, S6b, S6c) on release of 51Cr, production of inositol triphosphate (IP3), and release of arachidonic acid (AA) were examined in cultured microvascular endothelium derived from human brain (HBEC). ET-1 induced dose-dependent release of 51CR (EC50 = 7 +/- 2 nM), transient increase of IP3 (EC50 = 0.67 +/- 0.09 nM), and sustained release of AA (EC50= 59 +/- 7 nM) from HBEC. Under the same experimental conditions, viability of the cells was preserved (> 97%) as assessed by exclusion of vital dye trypan blue and release of lactate dehydrogenase (LDH). Dexamethasone (1 microM) inhibited ET-1-induced AA release, whereas it was ineffective on 51Cr release. Protein kinase C (PKC) inhibitor H7 (200 nM), calcium channel blocker verapamil (10 microM), or IP3 receptor antagonist ryonidine (5 microM) reduced ET-1 (100 nM)-induced release of 51Cr. These findings indicate that endothelins can induce an increase of HBEC permeability by a receptor-specific activation of PKC and intracellular calcium mobilization.
Stanimirovic
D B
DB
Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
McCarron
R
R
Bertrand
N
N
Spatz
M
M
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D016718:Arachidonic Acid / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002560:Cerebrovascular Circulation; D002860:Chromium Radioisotopes; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D016232:Endothelins / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008833:Microcirculation; D010879:Piperazines / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D012433:Ryanodine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D002860:Chromium Radioisotopes; D016232:Endothelins; D007546:Isoquinolines; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D012433:Ryanodine; D016718:Arachidonic Acid; D003907:Dexamethasone; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D015544:Inositol 1,4,5-Trisphosphate; D014700:Verapamil; D007770:L-Lactate Dehydrogenase; D013755:Tetradecanoylphorbol Acetate