关键词: 15-酮基二十碳四烯酸, 肺动脉平滑肌细胞, 细胞内Ca2 + 浓度

Abstract: Abstract: Objective To explore the effect and source of 15-KETE on the cytosolic ［Ca^{2 +}］i in rat pulmonary arterial smooth cells( PASMCs) ． Method PASMCs obtained with enzyme ( collagenase I plus elastase) dispersing method were cultured in the first passage in vitro． Cells with concentration of 2 × 10 ^{－ 5} cells /mL were seeded onto a glass coverslip in 6 well plate，and cultured for 24 h in the incubator at 37℃． After cell adherence，the glass coverslip was taken out from 6 well plate，placed in special-made chamber，and washed with D-Hanks solution for 3 times． Cells were loaded with Fluo-3 /AM ( final concentration 10μmol /L) ，followed in the incubator at 37℃ for 30 min，then washed with D-Hanks solution for 3 times，extracellular dye was washed out，the Fluo-3 /AM loaded cells were available for ［Ca^{2 +}］i measurement within 2h． Using laser-scanning confocal microscope technique，the effect of 15-KETE on cytosolic ［Ca^{2 +}］i in rat PASMCs was measured． Ｒesult ( 1) 15-KETE ( 1 × 10 － 8 ～ 1 × 10 － 6 mol /L) increased cytosolic ［Ca^{2 +}］i of rat PASMCs in a concentration-dependent manner． ( 2) The L-type Ca^{2 +} channel blocker verapamil ( 10 － 6 mol /L) and Ca^{2 +} -free Krebs solution significantly inhibited the increase of cytosolic ［Ca^{2 +}］i induced by 15-KETE． Conclusion 15-KETE may increase cytosolic ［Ca^{2 +}］i of rat PASMCs， and the source of Ca^{2 +} was from intercellular solution．

Key words: 15-ketoeicosatetretraenoic acid, pulmonary arterial smooth cells, ［Ca^{2 +}］i