Natsuki R
Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata, Japan.
Arukoru Kenkyuto Yakubutsu Ison. 1995 Oct;30(5):348-57.
The activities of phospholipase A2 (PLA2) and C (PLC) in subcellulal fractions from chick embryo brain, heart and liver were determined using the substrate 1-palmitoy 1-2-N-(4-nitrobenzo-2-oxa-1, 3-diazole amino caproyl-phsphatidylcholine (NBD-PC), and the effect of chronic ethanol treatment on these activities was evaluated. PLA2 and PLC activities of each fraction were assayed by measuring release of N- (4-nitrobenzo-2-oxa-1, 3-diazole) amino caproic acid (NBD-caproic acid) and of 1-palmitoy 1-2-NBD amino caproyl glycerol (NBD-DG) from exogenous NBD-PC. The microsomal membrane fluidity was estimated from diphenylhexatriene anisotropy (gamma). Cytosolic, mitochondorial, and microsomal subcellular fractions were prepared by differential centrifugation of homogenates of the brain, heart, and liver. Microsomal subcellular fractions from the brain, heart and liver of ethanol treated chick embryo showed significantly higher PLA2 and PLC specific activities than did corresponding fractions from non-treated chick embryo. Mitochondrial subcellular fractions from the brain and heart of ethanol-treated chick embryo also showed significantly higher PLA2 and PLC specific activities than the corresponding control fractions. Microsomal fractions from the brain and heart of ethanol-treated chick embryo decreased significantly the gamma than those of control. These results suggest that the change in the membrane fluidity is an apparent prerequisite for the changes of PLA2 and PLC activities.
使用底物1-棕榈酰基-2-N-(4-硝基苯并-2-恶唑-1,3-二唑氨基己酰基)磷脂酰胆碱(NBD-PC)测定了鸡胚脑、心脏和肝脏亚细胞组分中磷脂酶A2(PLA2)和磷脂酶C(PLC)的活性,并评估了慢性乙醇处理对这些活性的影响。通过测量从外源性NBD-PC释放的N-(4-硝基苯并-2-恶唑-1,3-二唑)氨基己酸(NBD-己酸)和1-棕榈酰基-2-NBD氨基己酰基甘油(NBD-DG)来测定各组分的PLA2和PLC活性。通过二苯基己三烯各向异性(γ)估计微粒体膜流动性。通过对脑、心脏和肝脏匀浆进行差速离心制备胞质、线粒体和微粒体亚细胞组分。乙醇处理的鸡胚脑、心脏和肝脏的微粒体亚细胞组分显示出比未处理鸡胚的相应组分显著更高的PLA2和PLC比活性。乙醇处理的鸡胚脑和心脏的线粒体亚细胞组分也显示出比相应对照组分显著更高的PLA2和PLC比活性。乙醇处理的鸡胚脑和心脏的微粒体组分的γ比对照组显著降低。这些结果表明膜流动性的变化是PLA2和PLC活性变化的明显前提条件。