Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease.
Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease.
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DOI：
10.1016/j.redox.2017.08.020
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发表时间：
2018-04
期刊：
影响因子：
11.4
通讯作者：
Kelm M
中科院分区：
文献类型：
--
作者：
Cortese-Krott MM;Mergia E;Kramer CM;Lückstädt W;Yang J;Wolff G;Panknin C;Bracht T;Sitek B;Pernow J;Stasch JP;Feelisch M;Koesling D;Kelm M
内皮功能障碍与NO生物利用度降低和血管和血小板中NO受体可溶性鸟苷环化酶(SGC)的激活受损有关。红细胞(RBC)在低氧和常氧条件下产生NO，但sGC及其下游信号通路包括磷酸二酯酶(PDE)-5和蛋白激酶G(PKG)在红细胞中表达和/或活性的证据仍有争议。在本研究中，我们旨在研究红细胞是否携带有功能的sGC信号通路，并探讨该通路在冠状动脉疾病(CAD)中是否受损。在此，我们使用两个独立的层析程序，证明了人和小鼠红细胞携带有催化活性的α1β1-sGC(异构体1)，它能将32P-GTP转化为32P-cGMP，以及PDE5和PKG。NO+Bay 41-2272对sGC的特异性刺激使细胞内cGMP-水平增加1000倍，并伴随着典型的PKG/VASP信号通路的激活。这种对NO的反应在α1-sGC基因敲除(KO)红细胞中被钝化，但在α2-sGC KO中完全保留。在稳定性冠心病和内皮功能障碍患者中，与老年对照组相比，红细胞eNOS表达降低；相反，红细胞sGC表达/活性和对NO的反应性完全保留，尽管两组sGC氧化增加。总之，我们的数据表明，在内皮功能障碍患者的红细胞中存在完整的sGC/PDE5/PKG依赖的信号通路，该通路对NO和sGC刺激物/激活剂仍有完全的反应。以此通路为靶点可能有助于治疗微循环中没有缺陷的疾病，如镰状细胞性贫血、肺动脉高压和心力衰竭。人和小鼠红细胞携带具有催化活性的α1蛋白1-糖蛋白(β1-sGC)、蛋白激酶G和蛋白水解酶5。NO刺激以sGC和PDE依赖的方式增加完整红细胞的cGMP-水平，NO刺激诱导典型的PKG/VASP依赖的信号转导。红细胞sGC的反应性在α1-sGC KO小鼠中没有被钝化，但在α2-sGC KO和eNOS KO小鼠中被保存。稳定性冠状动脉疾病患者的红细胞sGC活性完全保留。
Endothelial dysfunction is associated with decreased NO bioavailability and impaired activation of the NO receptor soluble guanylate cyclase (sGC) in the vasculature and in platelets. Red blood cells (RBCs) are known to produce NO under hypoxic and normoxic conditions; however evidence of expression and/or activity of sGC and downstream signaling pathway including phopshodiesterase (PDE)-5 and protein kinase G (PKG) in RBCs is still controversial. In the present study, we aimed to investigate whether RBCs carry a functional sGC signaling pathway and to address whether this pathway is compromised in coronary artery disease (CAD). Using two independent chromatographic procedures, we here demonstrate that human and murine RBCs carry a catalytically active α1β1-sGC (isoform 1), which converts 32P-GTP into 32P-cGMP, as well as PDE5 and PKG. Specific sGC stimulation by NO+BAY 41-2272 increases intracellular cGMP-levels up to 1000-fold with concomitant activation of the canonical PKG/VASP-signaling pathway. This response to NO is blunted in α1-sGC knockout (KO) RBCs, but fully preserved in α2-sGC KO. In patients with stable CAD and endothelial dysfunction red cell eNOS expression is decreased as compared to aged-matched controls; by contrast, red cell sGC expression/activity and responsiveness to NO are fully preserved, although sGC oxidation is increased in both groups. Collectively, our data demonstrate that an intact sGC/PDE5/PKG-dependent signaling pathway exists in RBCs, which remains fully responsive to NO and sGC stimulators/activators in patients with endothelial dysfunction. Targeting this pathway may be helpful in diseases with NO deficiency in the microcirculation like sickle cell anemia, pulmonary hypertension, and heart failure. Human and murine RBCs carry catalytically active α1β1-sGC (isoform 1), PKG and PDE5. NO-stimulation increases cGMP-levels in intact RBCs in a sGC and PDE-dependent manner, NO stimulation induces canonical PKG/VASP-dependent signaling. NO responsiveness of red cell sGC is blunted in α1-sGC KO mice, but is preserved in α2-sGC KO and eNOS KO mice. Red cell sGC activity is fully preserved in patients with stable coronary artery disease.
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影响因子：
3.7
作者：
Eligini S;Porro B;Lualdi A;Squellerio I;Veglia F;Chiorino E;Crisci M;Garlaschè A;Giovannardi M;Werba JP;Tremoli E;Cavalca V
通讯作者：
Cavalca V
影响因子：
16.6
作者：
Follmann, Markus;Griebenow, Nils;Straub, Alexander
通讯作者：
Straub, Alexander
影响因子：
7.4
作者：
Erkens, Ralf;Kramer, Christian M.;Cortese-Krott, Miriam M.
通讯作者：
Cortese-Krott, Miriam M.
影响因子：
82.9
作者：
Cosby, K;Partovi, KS;Gladwin, MT
通讯作者：
Gladwin, MT
影响因子：
11.4
作者：
Cortese-Krott MM;Kelm M
通讯作者：
Kelm M