Activating P2Y1 receptors improves function in arteries with repressed autophagy
Activating P2Y1 receptors improves function in arteries with repressed autophagy
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DOI：
10.1093/cvr/cvac061
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发表时间：
2022-04-14
影响因子：
10.8
通讯作者：
Symons, J. David
中科院分区：
文献类型：
--
作者：
Cho, Jae Min;Park, Seul-Ki;Symons, J. David
目的在健康和疾病的背景下，内皮细胞(EC)自噬对血管内稳态的重要性正在演变。早些时候，我们报道了完整的EC自噬是维持切应力诱导的一氧化氮(NO)产生所必需的，这是通过糖酵解依赖的嘌呤能信号传递给内皮NO合成酶(ENOS)所必需的。在这里，我们说明了这些发现的翻译和功能意义。方法和结果首先，与成年对照组相比，我们使用表现出钝化的EC自噬和动脉功能受损的老年男性人类和小鼠来评估翻译相关性。在成人和老年受试者中，由有节奏的握力运动引起的活动性充血--与基线相似的径动脉切变率升高，持续60分钟。与基线相比，成人而不是老年志愿者的桡动脉内皮细胞出现了自噬启动、p-eNOS(S1177)激活和NO生成的指标。对于小鼠，自噬和p-eNOS(S1177)激活的指数在成年动物的内皮细胞中表现强劲，但在完成60分钟跑步机跑步的老年动物中则不是。此外，20dyne中心点cm(2)层流切应力x 45min可增加成年小鼠动脉内皮细胞的自噬流量、糖酵解、ATP生成和p-eNOS(S1177)。关于功能相关性，我们接下来质疑体外无法启动EC自噬、糖酵解和p-eNOS(S1177)是否会导致体外动脉功能障碍。在成年小鼠的血管中，(I)没有合酶抑制；(Ii)使用3-甲基腺嘌呤(3-MA)的急性自噬损伤；(Iii)EC Atg3耗竭(iecAtg3KO小鼠)；(Iv)嘌呤能2Y(1)受体(P2Y(1)-R)阻断；以及(V)胚系P2Y(1)-Rs的耗竭，在成年小鼠的血管中显示了受损的管腔内血流介导的血管扩张。重要的是，使用2-甲硫基-ADP(2-Me-ADP)激活的P2Y(1)-R改善了来自(I)用3-MA治疗的成年小鼠；(Ii)成年iecAtg3KO小鼠；以及(Iii)有抑制EC自噬的老年动物的动脉扩张能力。结论通过激活P2Y(1)-Rs，可改善伴有药物、遗传和年龄相关的EC自噬损害的动脉功能障碍。
Aim The importance of endothelial cell (EC) autophagy to vascular homeostasis in the context of health and disease is evolving. Earlier, we reported that intact EC autophagy is requisite to maintain shear-stress-induced nitric oxide (NO) generation via glycolysis-dependent purinergic signalling to endothelial NO synthase (eNOS). Here, we illustrate the translational and functional significance of these findings. Methods and results First, we assessed translational relevance using older male humans and mice that exhibit blunted EC autophagy and impaired arterial function vs. adult controls. Active hyperaemia evoked by rhythmic handgrip exercise-elevated radial artery shear-rate similarly from baseline in adult and older subjects for 60 min. Compared with baseline, indexes of autophagy initiation, p-eNOS(S1177) activation, and NO generation, occurred in radial artery ECs obtained from adult but not older volunteers. Regarding mice, indexes of autophagy and p-eNOS(S1177) activation were robust in ECs from adult but not older animals that completed 60-min treadmill-running. Furthermore, 20 dyne center dot cm(2) laminar shear stress x 45-min increased autophagic flux, glycolysis, ATP production, and p-eNOS(S1177) in primary arterial ECs obtained from adult but not older mice. Concerning functional relevance, we next questioned whether the inability to initiate EC autophagy, glycolysis, and p-eNOS(S1177) in vitro precipitates arterial dysfunction ex vivo. Compromised intraluminal flow-mediated vasodilation displayed by arteries from older vs. adult mice was recapitulated in vessels from adult mice by (i) NO synthase inhibition; (ii) acute autophagy impairment using 3-methyladenine (3-MA); (iii) EC Atg3 depletion (iecAtg3KO mice); (iv) purinergic 2Y(1)-receptor (P2Y(1)-R) blockade; and (v) germline depletion of P2Y(1)-Rs. Importantly, P2Y(1)-R activation using 2-methylthio-ADP (2-Me-ADP) improved vasodilatory capacity in arteries from (i) adult mice treated with 3-MA; (ii) adult iecAtg3KO mice; and (iii) older animals with repressed EC autophagy. Conclusions Arterial dysfunction concurrent with pharmacological, genetic, and age-associated EC autophagy compromise is improved by activating P2Y(1)-Rs.