Maghni K, Robidoux C, Laporte J, Hallée A, Sirois P
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, PQ, Canada.
Prostaglandins. 1990 Aug;40(2):217-27. doi: 10.1016/0090-6980(90)90085-a.
Lung cells have been isolated by enzymatic digestion of guinea pig lungs and mechanical dispersion to obtain a suspension of viable cells (approximately 500 X 10(6) cells). Type II pneumocytes have been purified to approximately 92% by centrifugal elutriation (2000 rpm, 15 ml/min) followed by a plating in plastic dishes coated with guinea pig IgG (500 micrograms/ml). We have investigated the arachidonic acid metabolism through the cyclooxygenase pathway in this freshly isolated type II cells (2 x 10(6) cells/ml). Purified type II pneumocytes produced thromboxane B2 (TxB2) predominantly and to a smaller extent the 6-keto prostaglandin PGF1 alpha (6-keto-PGF1 alpha) and prostaglandin E2 (PGE2) after incubation with 10 microM arachidonic acid. The stimulation of pneumocytes with 2 microM calcium ionophore A23187 released less eicosanoids than were produced when cells were incubated with 10 microM arachidonic acid. There was no additive effect when the cells were treated with both arachidonic acid and the ionophore A23187. Guinea pig type II pneumocytes failed to release significant amounts of TxB2, 6-keto-PGF1 alpha and PGE2 after stimulation with 10 nM leukotriene B4, 10 nM leukotriene D4, 10 nM platelet-activating factor, 5 microM formyl-methionyl-leucyl-phenylalanine, 0.2 microM bradykinin and 10 nM phorbol myristate acetate. Our findings indicate that guinea pig type II pneunomocytes possess the enzymatic machinery necessary to convert arachidonic acid to specific cyclooxygenase products, which may suggest a role for these cells in lung inflammatory processes.
通过酶消化豚鼠肺脏并进行机械分散,已分离出肺细胞,以获得活细胞悬液(约500×10⁶个细胞)。通过离心淘析(2000转/分钟,15毫升/分钟),随后接种于涂有豚鼠IgG(500微克/毫升)的塑料培养皿中,II型肺细胞已被纯化至约92%。我们研究了这种新鲜分离的II型细胞(2×10⁶个细胞/毫升)中通过环氧化酶途径的花生四烯酸代谢。纯化的II型肺细胞在与10微摩尔花生四烯酸孵育后,主要产生血栓素B2(TxB2),少量产生6-酮前列腺素PGF1α(6-酮-PGF1α)和前列腺素E2(PGE2)。用2微摩尔钙离子载体A23187刺激肺细胞释放的类花生酸比用10微摩尔花生四烯酸孵育细胞时产生的少。当细胞同时用花生四烯酸和离子载体A23187处理时,没有相加效应。在用10纳摩尔白三烯B4、10纳摩尔白三烯D4、10纳摩尔血小板活化因子、5微摩尔甲酰甲硫氨酰亮氨酰苯丙氨酸、0.2微摩尔缓激肽和10纳摩尔佛波醇肉豆蔻酸酯刺激后,豚鼠II型肺细胞未能释放大量的TxB2、6-酮-PGF1α和PGE2。我们的研究结果表明,豚鼠II型肺细胞拥有将花生四烯酸转化为特定环氧化酶产物所需的酶机制,这可能表明这些细胞在肺部炎症过程中发挥作用。