Maghni K, Robidoux C, Laporte J, Hallee A, Borgeat P, Sirois P
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, P.Q., Canada.
Prostaglandins. 1991 Sep;42(3):251-67. doi: 10.1016/0090-6980(91)90114-u.
Pulmonary and splenic Kurloff cells have been purified from estrogen-treated guinea pig. Enzymatic digestion of lung tissue and mechanical dispersion of cells yielded about 650 x 10(6) viable cells. After centrifugal elutriation and centrifugation on continuous Percoll gradient, a population of high-density (1,100 g/ml) pulmonary Kurloff cells were obtained with high viability (approximately 99%) and purity (approximately 99%). Splenic Kurloff cells have been isolated by disruption of spleen tissue and centrifugation on continuous Percoll gradient. High-density splenic Kurloff cells (150 x 10(6) cells per spleen) were also obtained with high purity (approximately 99%) and viability (approximately 99%). Pulmonary and splenic Kurloff cells were incubated with various concentrations of arachidonic acid (10, 30 and 100 microM) in the absence or presence of 2 microM ionophore A23187. With 10 microM arachidonic acid the relative production of cyclooxygenase products was the following: TxB2 greater than PGE2 approximately PGI2. For an arachidonic acid concentration superior to 10 microM, the profile of release was PGE2 much greater than TxB2 greater than PGI2. Arachidonic acid metabolism through the 5-lipoxygenase pathway was also studied by incubating pulmonary or splenic Kurloff cells with 10 microM arachidonic acid in the absence or presence of 2 microM ionophore A23187, or in some experiments, with 2.5 microM leukotriene A4. Reverse phase HPLC profiles clearly indicated that high-density Kurloff cells did not express 5-lipoxygenase activity. However, these cells showed the ability to convert exogenous leukotriene A4 into leukotriene B4 suggesting the presence of LTA4 hydrolase activity. These data have been confirmed by a sensitive RIA method. This study constitutes the first report on the purification of pulmonary Kurloff cells and on arachidonic acid metabolism by these cells. The possible implications of Kurloff cells in various biological events are discussed.
已从经雌激素处理的豚鼠中纯化出肺和脾的库尔洛夫细胞。对肺组织进行酶消化并机械分散细胞，得到约650×10⁶个活细胞。经过离心淘析和在连续Percoll梯度上离心后，获得了高密度（1100 g/ml）的肺库尔洛夫细胞群体，其活力高（约99%）且纯度高（约99%）。通过破坏脾组织并在连续Percoll梯度上离心来分离脾库尔洛夫细胞。也获得了高密度的脾库尔洛夫细胞（每个脾150×10⁶个细胞），其纯度高（约99%）且活力高（约99%）。在不存在或存在2 μM离子载体A23187的情况下，将肺和脾的库尔洛夫细胞与不同浓度的花生四烯酸（10、30和100 μM）一起孵育。对于10 μM的花生四烯酸，环氧化酶产物的相对产生情况如下：TXB₂＞PGE₂≈PGI₂。对于高于10 μM的花生四烯酸浓度，释放情况为PGE₂远大于TXB₂＞PGI₂。还通过在不存在或存在2 μM离子载体A23187的情况下，将肺或脾的库尔洛夫细胞与10 μM花生四烯酸一起孵育，或在一些实验中与2.5 μM白三烯A4一起孵育，研究了通过5-脂氧合酶途径的花生四烯酸代谢。反相高效液相色谱图谱清楚地表明，高密度库尔洛夫细胞不表达5-脂氧合酶活性。然而，这些细胞显示出将外源性白三烯A4转化为白三烯B4的能力，这表明存在LTA4水解酶活性。这些数据已通过灵敏的放射免疫分析方法得到证实。本研究构成了关于肺库尔洛夫细胞纯化以及这些细胞花生四烯酸代谢的首篇报道。讨论了库尔洛夫细胞在各种生物学事件中的可能意义。
