Vigne P, Hechler B, Gachet C, Breittmayer J P, Frelin C
Institut de Pharmacologie Moléculaire et Cellulaire, CNRS UPR 411, 660 Route des Lucioles, Valbonne, 06560, France.
Biochem Biophys Res Commun. 1999 Mar 5;256(1):94-7. doi: 10.1006/bbrc.1999.9558.
The effects of 2'- and 3'-O-(4-benzoylbenzoyl)-ATP (BzATP) on intracellular Ca2+ mobilization and cyclic AMP accumulation were investigated using rat brain capillary endothelial cells which express an endogenous P2Y1 receptor, human platelets which are known to express a P2Y1 receptor, and Jurkat cells stably transfected with the human P2Y1 receptor. In endothelial cells, BzATP was a competitive inhibitor of 2-methylthio ADP (2-MeSADP) and ADP induced [Ca2+]i responses (Ki = 4.7 microM) and reversed the inhibition by ADP of adenylyl cyclase (Ki = 13 microM). In human platelets, BzATP inhibited ADP-induced aggregation (Ki = 5 microM), mobilization of intracellular Ca2+ stores (Ki = 6.3 microM), and inhibition of adenylyl cyclase. In P2Y1-Jurkat cells, BzATP inhibited ADP and 2-MeSADP-induced [Ca2+]i responses (Ki = 2.5 microM). It was concluded that BzATP is an antagonist of rat and human P2Y1 receptors and of platelet aggregation. In contrast to other P2Y1 receptor antagonists (A2P5P and A3P5P) which inhibit only ADP-induced Ca2+ mobilization, BzATP inhibits both the Ca2+- and the cAMP-dependent intracellular signaling pathways of ADP. These results provide further evidence that P2Y1 receptors contribute to platelet ADP responses.
使用表达内源性P2Y1受体的大鼠脑微血管内皮细胞、已知表达P2Y1受体的人血小板以及稳定转染人P2Y1受体的Jurkat细胞，研究了2'-和3'-O-(4-苯甲酰苯甲酰基)-ATP（BzATP）对细胞内Ca2+动员和环磷酸腺苷（cAMP）积累的影响。在内皮细胞中，BzATP是2-甲硫基ADP（2-MeSADP）和ADP诱导的[Ca2+]i反应的竞争性抑制剂（Ki = 4.7 microM），并逆转了ADP对腺苷酸环化酶的抑制作用（Ki = 13 microM）。在人血小板中，BzATP抑制ADP诱导的聚集（Ki = 5 microM）、细胞内Ca2+储存的动员（Ki = 6.3 microM）以及对腺苷酸环化酶的抑制作用。在P2Y1-Jurkat细胞中，BzATP抑制ADP和2-MeSADP诱导的[Ca2+]i反应（Ki = 2.5 microM）。得出的结论是，BzATP是大鼠和人P2Y1受体以及血小板聚集的拮抗剂。与仅抑制ADP诱导的Ca2+动员的其他P2Y1受体拮抗剂（A2P5P和A3P5P）不同，BzATP抑制ADP的Ca2+依赖性和cAMP依赖性细胞内信号通路。这些结果进一步证明P2Y1受体参与血小板ADP反应。
