Role of phosphodiesterase type 3A and 3B in regulating platelet and cardiac function using subtype-selective knockout mice.
Role of phosphodiesterase type 3A and 3B in regulating platelet and cardiac function using subtype-selective knockout mice.
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DOI：
10.1016/j.cellsig.2007.03.012
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发表时间：
2007-08
影响因子：
4.8
通讯作者：
Bing Sun;Haiquan Li;Y. Shakur;J. Hensley;S. Hockman;J. Kambayashi;V. Manganiello;Yongge Liu
中科院分区：
文献类型：
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作者：
Bing Sun;Haiquan Li;Y. Shakur;J. Hensley;S. Hockman;J. Kambayashi;V. Manganiello;Yongge Liu
磷酸二酯酶3型（PDE 3）是心血管系统内cAMP介导的反应的重要调节剂。PDE 3存在两种亚型：PDE 3A和PDE 3B，具有不同的细胞和亚细胞位置。由于缺乏亚型特异性药理学工具，尚未确定每种亚型在调节心血管功能中的确切作用。在这项研究中，我们研究了血小板和心脏功能，使用PDE 3A和PDE 3B基因敲除（KO）小鼠。从KO和年龄匹配的野生型（WT）小鼠的血液制备富血小板血浆。PGE 1（1 μg/mL）几乎完全抑制WT、PDE 3A KO和PDE 3B KO小鼠的血小板聚集。在WT小鼠的血小板中，选择性PDE 3抑制剂西洛酰胺（100 μM）可阻断胶原和ADP诱导的聚集。相比之下，西洛酰胺对PDE 3A KO小鼠的血小板聚集没有影响。在PDE 3B KO小鼠中，对胶原蛋白和ADP诱导的血小板聚集的抑制与WT小鼠相似。PDE 3A KO小鼠血小板中的静息血小板内cAMP浓度是WT血小板中的两倍。PGE 1（0.1 μg/mL）刺激后，PDE 3A KO小鼠血小板中的细胞内cAMP浓度比WT小鼠显著增加。在体内，PDE 3A KO小鼠可免受胶原蛋白/肾上腺素诱导的肺血栓形成和死亡，而在PDE 3B KO小鼠中未观察到此类保护。与年龄匹配的WT小鼠相比，PDE 3A KO小鼠的心率显著更高，而PDE 3B KO小鼠的心率与WT相似。PDE 3A或PDE 3B KO小鼠之间的心肌收缩力无差异。在两种类型的KO小鼠中，异丙肾上腺素以类似的剂量依赖性方式增加心率和收缩力。西洛酰胺增加WT和PDE 3B KO小鼠的心率和收缩力，但在PDE 3A KO小鼠中不增加。与WT和PDE 3B KO小鼠相比，从PDE 3A KO小鼠心脏制备的膜组分中的环AMP-PDE活性较低，且不受西洛酰胺抑制。PDE 3A是PDE 3在血小板和心室肌细胞中表达的主要亚型，并负责PDE 3抑制引起的功能变化。
Phosphodiesterase type 3 (PDE3) is an important regulator of cAMP-mediated responses within the cardiovascular system. PDE3 exists as two subtypes: PDE3A and PDE3B, with distinct cellular and subcellular locations. Due to the lack of subtype-specific pharmacological tools, the definitive role of each subtype in regulating cardiovascular function has not been determined. In this study, we investigated platelet and cardiac function, using PDE3A and PDE3B gene knockout (KO) mice. Platelet-rich-plasma was prepared from the blood of KO and age-matched wild-type (WT) mice. PGE1(1 μg/mL) almost completely inhibited aggregation of platelets from WT, PDE3A KO and PDE3B KO mice. In platelets from WT mice, cilostamide (100 μM), a selective PDE3 inhibitor, blocked collagen- and ADP-induced aggregation. In contrast, cilostamide had no effect on aggregation of platelets from PDE3A KO mice. In PDE3B KO mice, inhibition of collagen- and ADP-induced platelet aggregation was similar to that in WT mice. The resting intra-platelet cAMP concentration in platelets from PDE3A KO mice was twice that in the WT platelets. After PGE1(0.1 μg/mL) stimulation, intra-cellular cAMP concentration was increased significantly more in platelets from PDE3A KO mice compared to WT mice. In vivo, PDE3A KO mice were protected against collagen/epinephrine-induced pulmonary thrombosis and death, while no such protection was observed in PDE3B KO mice. The heart rate of PDE3A KO mice was significantly higher, compared with age-matched WT mice, while that of PDE3B KO mice was similar to WT. There was no difference in cardiac contractility between PDE3A or PDE3B KO mice. Heart rate and contractility were increased in a similar dose-dependent fashion by isoproterenol in both types of KO mice. Cilostamide increased heart rate and contractility in WT and PDE3B KO but not in PDE3A KO mice. Compared to WT and PDE3B KO mice, cyclic AMP-PDE activity in membrane fractions prepared from the hearts of PDE3A KO mice was lower and not inhibited by cilostamide. The data suggest that PDE3A is the main subtype of PDE3 expressed in platelets and cardiac ventricular myocytes, and is responsible for the functional changes caused by PDE3 inhibition.