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Elevated arterial shear rate increases indices of endothelial cell autophagy and nitric oxide synthase activation in humans.

Continuous laminar shear stress increases the process of autophagy, activates p-eNOSS1177 , and generates nitric oxide (NO) in bovine and human arterial endothelial cells (ECs) compared to static controls. However, the translational relevance of these findings has not been explored. In the current study, primary ECs were collected from the radial artery (RA) of 7 adult males using sterile j-wires before (Pre) and after (Post) 60-min of rhythmic handgrip exercise (HG) performed with the same arm. After ECs were identified by positive co-staining for VE-cadherin and DAPI, immunofluorescent antibodies were used to assess indices of autophagy, NO generation, and superoxide anion (O2•- ) production. Commercially available primary human arterial ECs were stained and processed in parallel to serve as controls. All endpoints were evaluated using 75 ECs from each subject. Relative to Pre-HG, HG elevated arterial shear rate (p<0.05) ~ 3-fold, while heart rate, arterial pressure, and cardiac output were not altered. Compared to values obtained from ECs Pre-HG, Post-HG ECs displayed increased (p<0.05) expression of p-eNOSS1177 , NO generation, O2•- production, BECLIN-1, microtubule-associated proteins 1A/1B light chain 3B (LC3B), autophagy-related gene 3 (ATG3), lysosomal associated membrane protein 2A (LAMP2A), and decreased (p<0.05) expression (i.e., enhanced degradation) of the adapter protein p62/SQSTM1. These novel findings provide evidence that elevated arterial shear rate associated with functional hyperemia initiates autophagy, activates p-eNOSS1177 , and increases NO and O2•- generation in primary human ECs.

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