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首页> 外文期刊>American Journal of Physiology >Purine and pyrimidine nucleotide-sensitive phospholipase A(2) in ampulla from frog semicircular canal.
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Purine and pyrimidine nucleotide-sensitive phospholipase A(2) in ampulla from frog semicircular canal.

机译：嘌呤和嘧啶核苷酸敏感的磷脂酶A（2）在青蛙半圆形管的壶腹中。

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摘要

This study was attempted to characterize pharmacologically the P2Y receptors triggering phospholipase A(2) (PLA(2)) activation in ampulla from frog semicircular canal. A microassay was developed to screen the abilities of UTP analogs to stimulate [(3)H]arachidonic acid release by labeled ampullas. At 26 degrees C UTP induced a dose-dependent and saturable increase of PLA(2) activity (apparent activation constant 1.3 +/- 0.4 microM, Hill coefficient 0.9 +/- 0.2, maximal stimulating factor 2.0 +/- 0.1). The rank order of potency of agonists for PLA(2) activation was UTP > or = UDP > adenosine 5'-O-(2-thiodiphosphate) = adenosine 5'-O-(3-thiotriphosphate) > or = ATP = 2-methylthio-ATP > or = ADP = diadenosine tetraphosphate > or = alpha,beta-methylene-ATP = CTP > 2' and 3'-O-(4-benzoylbenzoyl)-ATP > or = AMP = UMP uridine and adenosine. UTP- and 2-methylthio-ATP-induced PLA(2) activations were inhibited by U-73122, GF-109203X, and methyl arachidonyl fluorophosphate. Basal activity was stimulated by phorbol ester and epinephrine and reduced by vasotocin, isoproterenol, prostaglandin E(2), cAMP, and forskolin. H-89 restored the cAMP- and forskolin-inhibited PLA(2) activities. Results indicate that P2Y receptor-mediated PLA(2) stimulation requires phopholipase C and protein kinase C activations and basal activity is inhibited by agonist-stimulated cAMP-dependent mechanisms.
机译：这项研究试图从药理学上表征青蛙半圆形管壶腹中触发磷脂酶A（2）（PLA（2））活化的P2Y受体。开发了一种微测定法，以筛选UTP类似物刺激标记壶腹释放[（3）H]花生四烯酸的能力。在26摄氏度时，UTP诱导了PLA（2）活性的剂量依赖性和饱和增加（表观活化常数1.3 +/- 0.4 microM，希尔系数0.9 +/- 0.2，最大刺激因子2.0 +/- 0.1）。激动剂对PLA（2）激活的效力的排名顺序是UTP>或= UDP>腺苷5'-O-（2-硫代二磷酸）=腺苷5'-O-（3-硫代三磷酸）>或= ATP = 2-甲硫基ATP>或= ADP =二磷酸腺苷>或=α，β-亚甲基ATP = CTP> 2'和3'-O-（4-苯甲酰基苯甲酰基）-ATP>或= AMP = UMP 尿苷和腺苷。 UTP-和2-甲硫基ATP诱导的PLA（2）激活被U-73122，GF-109203X和甲基花生四烯基氟磷酸盐抑制。佛波酯和肾上腺素可刺激基础活性，血管收缩素，异丙肾上腺素，前列腺素E（2），cAMP和福司可林可降低基础活性。 H-89恢复了cAMP和毛喉素抑制的PLA（2）活动。结果表明，P2Y受体介导的PLA（2）刺激需要磷脂酶C和蛋白激酶C的激活，并且基础活性被激动剂刺激的cAMP依赖性机制抑制。

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