Phosphodiesterase 2 Mediates Redox-Sensitive Endothelial Cell Proliferation and Angiogenesis by Thrombin via Rac1 and NADPH Oxidase 2
Phosphodiesterase 2 Mediates Redox-Sensitive Endothelial Cell Proliferation and Angiogenesis by Thrombin via Rac1 and NADPH Oxidase 2
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DOI：
10.1161/circresaha.109.196592
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发表时间：
2009-05-22
影响因子：
20.1
通讯作者：
Goerlach, Agnes
中科院分区：
文献类型：
--
作者：
Diebold, Isabel;Djordjevic, Talija;Goerlach, Agnes
环核苷酸磷酸二酯酶（PDE）控制包括内皮细胞在内的许多细胞类型中的第二信使cAMP和cGMP的水平。虽然PDE 2具有被cGMP激活但水解cAMP的独特性质，但其在内皮功能中的作用知之甚少。活性氧（ROS）是一种重要的信号分子，控制着内皮细胞的多种功能。因此，我们研究了PDE 2是否与凝血酶诱导的人脐静脉内皮细胞的ROS生成和增殖反应有关。凝血酶刺激已知激活NADPH氧化酶的GTdR Rac 1，并增强ROS形成，而短发夹（sh）RNA抑制或耗尽PDE 2阻止了这些反应。用8-Br-cGMP或心房利钠肽进行了类似的观察。在协议中，凝血酶升高cGMP，但降低cAMP水平，而db-cAMP或毛喉素减少Rac 1活性和ROS的生产。随后，PDE 2过表达激活Rac 1，增加ROS生成，并增强增殖和体外毛细血管形成。在存在针对NADPH氧化酶亚基NOX 2的无活性Rac 1或shRNA的情况下，未观察到这些反应。PDE 2的抑制或耗竭也阻止凝血酶诱导的增殖和毛细血管形成。重要的是，通过慢病毒shRNA或PDE 2抑制下调PDE 2分别阻止了小鼠主动脉外植体的血管发芽和小鼠模型中的体内血管生成。总之，PDE 2促进NADPH氧化酶依赖性ROS产生的活化以及随后的内皮增殖和血管生成。靶向PDE 2可能为与内皮功能障碍、氧化应激、血管增殖和血管生成相关的疾病提供新的治疗方法。(Circ Res. 2009;104：1169-1177）。
Cyclic nucleotide phosphodiesterases (PDEs) control the levels of the second messengers cAMP and cGMP in many cell types including endothelial cells. Although PDE2 has the unique property to be activated by cGMP but to hydrolyze cAMP, its role in endothelial function is only poorly understood. Reactive oxygen species (ROS) have been recognized as signaling molecules controlling many endothelial functions. We thus investigated whether PDE2 would link to ROS generation and proliferative responses in human umbilical vein endothelial cells in response to thrombin. Thrombin stimulated the GTPase Rac1, known to activate NADPH oxidases, and enhanced ROS formation, whereas PDE2 inhibition or depletion by short hairpin (sh) RNA prevented these responses. Similar observations were made with 8-Br-cGMP or atrial natriuretic peptide. In agreement, thrombin elevated cGMP but decreased cAMP levels, whereas db-cAMP or forskolin diminished Rac1 activity and ROS production. Subsequently, PDE2 overexpression activated Rac1, increased ROS generation, and enhanced proliferation and in vitro capillary formation. These responses were not observed in the presence of inactive Rac1 or shRNA against the NADPH oxidase subunit NOX2. Inhibition or depletion of PDE2 also prevented thrombin-induced proliferation and capillary formation. Importantly, downregulation of PDE2 by lentiviral shRNA or PDE2 inhibition prevented vessel sprouting from mouse aortic explants and in vivo angiogenesis in a mouse model, respectively. In summary, PDE2 promotes activation of NADPH oxidase-dependent ROS production and subsequent endothelial proliferation and angiogenesis. Targeting PDE2 may provide a new therapeutic approach in diseases associated with endothelial dysfunction, oxidative stress, vascular proliferation, and angiogenesis. (Circ Res. 2009;104:1169-1177.)