Endothelial Cell Autophagy Maintains Shear Stress-Induced Nitric Oxide Generation via Glycolysis-Dependent Purinergic Signaling to Endothelial Nitric Oxide Synthase.
Endothelial Cell Autophagy Maintains Shear Stress-Induced Nitric Oxide Generation via Glycolysis-Dependent Purinergic Signaling to Endothelial Nitric Oxide Synthase.
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DOI：
10.1161/atvbaha.117.309510
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发表时间：
2017-09
期刊：
影响因子：
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通讯作者：
Symons JD
中科院分区：
文献类型：
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作者：
Bharath LP;Cho JM;Park SK;Ruan T;Li Y;Mueller R;Bean T;Reese V;Richardson RS;Cai J;Sargsyan A;Pires K;Anandh Babu PV;Boudina S;Graham TE;Symons JD
受损的内皮细胞自噬损害剪切应力诱导的一氧化氮（NO）的产生。我们确定了责任机制。在牛主动脉内皮细胞（EC）暴露于剪切应力的自噬妥协，减少葡萄糖摄取和EC糖酵解衰减ATP的生产。我们假设，减少糖酵解依赖性嘌呤能信号通过P2 Y-1受体，继发于受损的自噬在内皮细胞，防止剪切诱导的p-eNOS 1177和NO的产生。恢复葡萄糖转运和糖酵解的操作（例如，GLUT 1的过表达）或嘌呤能信号传导（例如，添加外源性ADP）拯救了具有受损自噬的EC中剪切诱导的p-eNOS 1177和NO产生。相反，通过GLUT 1 siRNA抑制葡萄糖转运，通过外核苷酸酶介导的ATP/ADP降解阻断嘌呤能信号传导（例如，腺苷三磷酸双磷酸酶），或使用药理学方法抑制P2 Y1受体（例如，MRS 2179）或遗传（例如，P2 Y1-R siRNA）程序，抑制具有完整自噬的EC中剪切诱导的p-eNOS 1177和NO产生。支持PKCδT505在将自噬依赖性嘌呤能介导的信号传递给eNOS中的中心作用，我们发现：（i）剪切应力诱导的PKCδT505活化磷酸化通过抑制自噬而被否定;（ii）剪切诱导的p-eNOS 1177和NO生成通过药理学（例如，苔藓抑素）或遗传（例如，CA-PKCδ）PKCδT505的活化和（iii）药理学（例如，罗特勒蛋白）和遗传（例如，PKCδ siRNA）PKCδ抑制可防止具有完整自噬的EC中剪切诱导的p-eNOSS 1177和NO的产生。在人动脉内皮细胞中揭示了自噬妥协后该通路中调节异常的关键节点。嘌呤能信号传导和/或PKCδ的靶向再活化具有恢复与抑制的EC自噬相关的病理中受损的NO产生的战略潜力。
Impaired endothelial cell autophagy compromises shear-stress induced nitric oxide (NO) generation. We determined the responsible mechanism. Upon autophagy compromise in bovine aortic endothelial cells (ECs) exposed to shear-stress, a decrease in glucose uptake and EC glycolysis attenuated ATP production. We hypothesized that decreased glycolysis-dependent purinergic signaling via P2Y-1 receptors, secondary to impaired autophagy in ECs, prevents shear-induced p-eNOSS1177 and NO generation. Maneuvers that restore glucose transport and glycolysis (e.g., overexpression of GLUT1) or purinergic signaling (e.g., addition of exogenous ADP) rescue shear-induced p-eNOSS1177 and NO production in ECs with impaired autophagy. Conversely, inhibiting glucose-transport via GLUT1 siRNA, blocking purinergic signaling via ectonucleotidase-mediated ATP/ADP degradation (e.g., apyrase), or inhibiting P2Y1 receptors using pharmacological (e.g., MRS2179) or genetic (e.g., P2Y1-R siRNA) procedures, inhibits shear-induced p-eNOSS1177 and NO generation in ECs with intact autophagy. Supporting a central role for PKCδT505 in relaying the autophagy-dependent purinergic-mediated signal to eNOS, we find that: (i) shear-stress induced activating phosphorylation of PKCδT505 is negated by inhibiting autophagy; (ii) shear-induced p-eNOSS1177 and NO generation are restored in autophagy-impaired ECs via pharmacological (e.g., bryostatin) or genetic (e.g., CA-PKCδ) activation of PKCδT505 and (iii) pharmacological (e.g., rottlerin) and genetic (e.g., PKCδ siRNA) PKCδ inhibition prevents shear-induced p-eNOSS1177 and NO generation in ECs with intact autophagy. Key nodes of dysregulation in this pathway upon autophagy compromise were revealed in human arterial endothelial cells. Targeted reactivation of purinergic signaling and/or PKCδ has strategic potential to restore compromised NO generation in pathologies associated with suppressed EC autophagy.