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Mechanotransduction is a process converting mechanical cues into electrochemical signals. Piezo1, a mechanically sensitive cation channel protein, is widely distributed in non-sensory cells of mammals, particularly highly expressed in the endothelial cells (ECs). Piezo1 is implicated in various physiological activities, including the regulation of vascular tone, angiogenesis, and maintenance of the endothelial barrier. Under pathological mechanical forces, Piezo1 is involved in the development of endothelial dysfunction-related diseases, including atherosclerosis (AS), hypertension, heart failure (HF), myocardial infarction (MI), pulmonary arterial hypertension (PAH), acute respiratory distress syndrome (ARDS), and diabetes mellitus (DM). In this review, we focus on the underlying mechanisms of Piezo1 in different endothelial dysfunction-related diseases, highlighting its roles in regulating endothelial function. Moreover, we present some activators and inhibitors targeting Piezo1, and discuss their different effects in distinct contexts. Overall, targeting Piezo1 may open a novel avenue of therapeutic tactic for endothelial dysfunction-related diseases.