Piezo1 channel activation stimulates ATP production through enhancing mitochondrial respiration and glycolysis in vascular endothelial cells

Background and purpose: Piezo1 channel is a mechanosensitive cationic channel which is activated by mechanical stretch or shear stress. Endothelial Piezo1 activation by shear stress of blood flow induces ATP release from endothelial cells, however, the link between the shear stress and endothelial ATP production is unclear.

Experimental approach: The mitochondrial respiratory function of cells was measured by using high-resolution respirometry system Oxygraph-2k. The intracellular Ca²⁺ concentration was evaluated by using Fluo-4/AM and mitochondrial Ca²⁺ concentration by Rhod-2/AM.

Key results: The specific Piezo1 channel activator Yoda1 or its analogue Dooku1 increased [Ca²⁺ ]_i in human umbilical vein endothelial cells (HUVECs), and both Yoda1 and Dooku1 increased mitochondrial oxygen consumption rates (OCRs) and mitochondrial ATP production in HUVECs and primary cultured rat aortic endothelial cells (RAECs). Knockdown of Piezo1 inhibited Yoda1 and Dooku1-induced increases of mitochondrial OCRs and mitochondrial ATP production in HUVECs. The shear stress mimics, Yoda1 and Dooku1, and the Piezo1 knock-down technique also demonstrated that Piezo1 activation increased glycolysis in HUVECs. Chelating extracellular Ca²⁺ with EGTA or chelating cytosolic Ca²⁺ with BAPTA-AM did not affect Yoda1 and Dooku1-induced increases of mitochondrial OCRs and ATP production, but chelating cytosolic Ca²⁺ inhibited Yoda1 and Dooku1-induced increase of glycolysis. Confocal microscopy showed that Piezo1 channel was present in mitochondria of endothelial cells, and Yoda1 and Dooku1 increased mitochondrial Ca²⁺ in endothelial cells.

Conclusion and implications: Piezo1 channel activation stimulates ATP production through enhancing mitochondrial respiration and glycolysis in vascular endothelial cells, suggesting a novel role of Piezo1 channel in endothelial ATP production.

Piezo1 activators; Piezo1 channel; endothelial cells; glycolysis; mitochondrial respiration.