Effect of calmodulin antagonists on lysosomal enzyme secretion and phospholipid metabolism in guinea-pig macrophages.
Effect of calmodulin antagonists on lysosomal enzyme secretion and phospholipid metabolism in guinea-pig macrophages.
复制标题
钙调蛋白拮抗剂对豚鼠巨噬细胞溶酶体酶分泌和磷脂代谢的影响。
DOI：
10.1042/bj2080549
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Y. Nagai
中科院分区：
文献类型：
--
作者：
T. Takenawa;Y. Homma;Y. Nagai
本文研究了钙调素拮抗剂对豚鼠腹腔巨噬细胞溶酶体酶分泌和脂质代谢的影响。钙调素拮抗剂，如三氟拉嗪，地布卡因和奎纳克林，抑制分泌N-乙酰-β-d-氨基葡萄糖苷酶从细胞松弛素B处理的巨噬细胞时，巨噬细胞被刺激的趋化肽，甲酰甲硫氨酰-亮氨酰-苯丙氨酸（f Met-Leu-Phe）或Ca（2+）离子载体A23187。研究了钙调素拮抗剂对[（32）P]P（i）或[（3）H]甘油掺入甘油脂以及[（14）C]甘油或[（3）H]花生四烯酸在[（14）C]甘油-或[（3）H]花生四烯酸-预标记脂质中再分布的影响。三氟拉嗪、地布卡因或奎纳克林刺激[（32）P]P（i）掺入磷脂酸（PtdA）和磷脂酰肌醇（PtdIns），而对磷脂酰乙醇胺（PtdEtn）、磷脂酰丝氨酸（PtdSer）、溶血磷脂酰胆碱（lyso-PtdCho）和溶血磷脂酰乙醇胺（lyso-PtdEtn）的标记无显著影响。而[（32）P]P（i）掺入磷脂酰胆碱（PtdCho）则受到抑制。当将钙调素拮抗剂加入到由fMet-Leu-Phe刺激的巨噬细胞中时，与仅由钙调素拮抗剂诱导的[（32）P]P（i）掺入PtdIns和PtdA相比，[（32）P]P（i）掺入协同增加。三氟拉嗪抑制[（3）H]甘油掺入PtdCho、三酰甘油和PtdEtn。同样在这种情况下，[（3）H]甘油掺入PtdA和PtdIns的能力大大增强。但对PtdSer、lyso-PtdEtn和lyso-PtdCho的[3 H]甘油掺入无影响。另一方面，用[（3）H]甘油标记的甘油二酯被10 μ m-三氟拉嗪最大程度地激活，并随着浓度的增加而稳定。当在脉冲追踪实验中检查钙调素拮抗剂对[（14）C]甘油在脂质中再分布的影响时，未检测到对PtdEtn、PtdCho、PtdIns、PtdSer、PtdA和三酰甘油和二酰甘油中[（14）C]甘油再分布的显著影响。用三氟拉嗪、地布卡因或奎纳克林处理巨噬细胞，[3] H]花生四烯酸标记的[3] H]花生四烯酸在PtdEtn和PtdCho中减少，而在PtdA中增加。[花生四烯酸-（3）H]二酰基甘油和非酯化[（3）H]-花生四烯酸的形成也得到增强，但[（3）H]花生四烯酸的增加仅在1和50 μ m之间的浓度下观察到。[花生四烯酸-（3）H]PtdIns未受到显著影响。在fMet-Leu-Phe存在下，这些药物对[花生四烯酸-（3）H]PtdA、二酰甘油和非酯化花生四烯酸的活化形成协同增强。
The effects of calmodulin antagonists on the secretion of lysosomal enzyme and lipid metabolism in guinea-pig peritoneal macrophages were studied. Calmodulin antagonists, such as trifluoperazine, dibucaine and quinacrine, inhibited the secretion of N-acetyl-beta-d-glucosaminidase from cytochalasin B-treated macrophages when the macrophages were stimulated by the chemotactic peptide, formylmethionyl-leucyl-phenylalanine (f Met-Leu-Phe) or the Ca(2+) ionophore A23187. The effect of calmodulin antagonists on the incorporation of [(32)P]P(i) or [(3)H]glycerol into glycerolipids as well as on the redistribution of [(14)C]glycerol or [(3)H]arachidonic acid in [(14)C]glycerol- or [(3)H]arachidonic acid-prelabelled lipids were examined. Trifluoperazine, dibucaine or quinacrine stimulated [(32)P]P(i) incorporation into phosphatidic acid (PtdA) and phosphatidylinositol (PtdIns) without significant effect on the labelling of phosphatidylethanolamine (PtdEtn), phosphatidylserine (PtdSer), lysophosphatidylcholine (lyso-PtdCho) and lysophosphatidylethanolamine (lyso-PtdEtn). The incorporation of [(32)P]P(i) into phosphatidylcholine (PtdCho) was, on the contrary, inhibited. When calmodulin antagonists were added to macrophages stimulated by fMet-Leu-Phe, [(32)P]P(i) incorporation into PtdIns and PtdA was synergistically increased compared with that induced only by calmodulin antagonists. Trifluoperazine inhibited the incorporation of [(3)H]glycerol into PtdCho, triacylglycerol and PtdEtn. Also in this case, the incorporation of [(3)H]glycerol into PtdA and PtdIns was greatly enhanced. But [(3)H]glycerol incorporation into PtdSer, lyso-PtdEtn and lyso-PtdCho was not affected by the drug. On the other hand, diacylglycerol labelling with [(3)H]glycerol was maximally activated by 10mum-trifluoperazine and levelled off with the increasing concentration. When the effect of calmodulin antagonists on the redistribution of [(14)C]glycerol among lipids was examined in pulse-chase experiments, no significant effect on [(14)C]glycerol redistribution in PtdEtn, PtdCho, PtdIns, PtdSer, PtdA and tri- and di-acylglycerol could be detected. When macrophages prelabelled with [(3)H]arachidonic acid were treated with trifluoperazine, dibucaine or quinacrine, the [(3)H]arachidonic acid moiety in PtdEtn and PtdCho was decreased and that in PtdA was increased. The formation of [arachidonate-(3)H]diacylglycerol and non-esterified [(3)H]-arachidonic acid was also enhanced, but the increase in [(3)H]arachidonic acid was only observed at concentrations between 1 and 50mum. [Arachidonate-(3)H]PtdIns was not significantly affected. The activated formation of [arachidonate-(3)H]PtdA, diacylglycerol and non-esterified arachidonic acid by these drugs was synergistically enhanced in the presence of fMet-Leu-Phe.