Endothelial Cell Autophagy Maintains Shear Stress-Induced Nitric Oxide Generation via Glycolysis-Dependent Purinergic Signaling to Endothelial Nitric Oxide Synthase
| Autor: | Seul Ki Park, Van Reese, Ting Ruan, Timothy E. Graham, Youyou Li, David D. Symons, Sihem Boudina, Jae Min Cho, Russel S. Richardson, Pon Velayutham Anandh Babu, Ashot Sargsyan, Leena P. Bharath, Robert A. Mueller, Tyler Bean, Jinjin Cai, Karla Maria Pereira Pires |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Nitric Oxide Synthase Type III Autophagy-Related Proteins Biology Nitric Oxide Transfection Mechanotransduction Cellular Article Nitric oxide 03 medical and health sciences chemistry.chemical_compound Receptors Purinergic P2Y1 Adenosine Triphosphate Shear stress Autophagy Serine Animals Humans Glycolysis Phosphorylation Protein Kinase Inhibitors Cells Cultured chemistry.chemical_classification Mice Knockout Reactive oxygen species Glucose Transporter Type 1 Endothelial nitric oxide synthase Endothelial Cells Purinergic signalling Cell biology Endothelial stem cell Mice Inbred C57BL Protein Kinase C-delta 030104 developmental biology chemistry Ubiquitin-Conjugating Enzymes Purinergic P2Y Receptor Antagonists Cattle RNA Interference Stress Mechanical Cardiology and Cardiovascular Medicine Reactive Oxygen Species |
| Zdroj: | Arteriosclerosis, thrombosis, and vascular biology. 37(9) |
| ISSN: | 1524-4636 |
| Popis: | Objective— Impaired endothelial cell (EC) autophagy compromises shear stress–induced nitric oxide (NO) generation. We determined the responsible mechanism. Approach and Results— On autophagy compromise in bovine aortic 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 P2Y1 (P2Y purinoceptor 1) receptors, secondary to impaired autophagy in ECs, prevents shear-induced phosphorylation of eNOS (endothelial nitric oxide synthase) at its positive regulatory site S1117 (p-eNOS S1177 ) and NO generation. Maneuvers that restore glucose transport and glycolysis (eg, overexpression of GLUT1 [glucose transporter 1]) or purinergic signaling (eg, addition of exogenous ADP) rescue shear-induced p-eNOS S1177 and NO production in ECs with impaired autophagy. Conversely, inhibiting glucose transport via GLUT1 small interfering RNA, blocking purinergic signaling via ectonucleotidase-mediated ATP/ADP degradation (eg, apyrase), or inhibiting P2Y1 receptors using pharmacological (eg, MRS2179 [2′-deoxy- N 6 -methyladenosine 3′,5′-bisphosphate tetrasodium salt]) or genetic (eg, P2Y1-receptor small interfering RNA) procedures inhibit shear-induced p-eNOS S1177 and NO generation in ECs with intact autophagy. Supporting a central role for PKCδ T505 (protein kinase C delta T505) in relaying the autophagy-dependent purinergic-mediated signal to eNOS, we find that (1) shear stress–induced activating phosphorylation of PKCδ T505 is negated by inhibiting autophagy, (2) shear-induced p-eNOS S1177 and NO generation are restored in autophagy-impaired ECs via pharmacological (eg, bryostatin) or genetic (eg, constitutively active PKCδ) activation of PKCδ T505 , and (3) pharmacological (eg, rottlerin) and genetic (eg, PKCδ small interfering RNA) PKCδ inhibition prevents shear-induced p-eNOS S1177 and NO generation in ECs with intact autophagy. Key nodes of dysregulation in this pathway on autophagy compromise were revealed in human arterial ECs. Conclusions— Targeted reactivation of purinergic signaling and PKCδ has strategic potential to restore compromised NO generation in pathologies associated with suppressed EC autophagy. |
| Databáze: | OpenAIRE |
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