Suppression of transient receptor potential melastatin 4 expression promotes conversion of endothelial cells into fibroblasts via transforming growth factor/activin receptor-like kinase 5 pathway

Objective: To study whether transient receptor potential melastatin 4 (TRPM4) participates in endothelial fibrosis and to investigate the underlying mechanism. Methods: Primary human endothelial cells were used and pharmacological and short interfering RNA-based approaches were used to test the transforming growth factor beta (TGF-beta)/activin receptor-like kinase 5 (ALK5) pathway participation and contribution of TRPM7 ion channel. Results: Suppression of TRPM4 expression leads to decreased endothelial protein expression and increased expression of fibrotic and extracellular matrix markers. Furthermore, TRPM4 downregulation increases intracellular Ca2+ levels as a potential condition for fibrosis. The underlying mechanism of endothelial fibrosis shows that inhibition of TRPM4 expression induces TGF-beta 1 and TGF-beta 2 expression, which act through their receptor, ALK5, and the nuclear translocation of the profibrotic transcription factor smad4. Conclusion: TRPM4 acts to maintain endothelial features and its loss promotes fibrotic conversion via TGF-beta production. The regulation of TRPM4 levels could be a target for preserving endothelial function during inflammatory diseases.