The potential of natural dietary polyphenols in the treatment of vascular diseases originating from veins has been suggested in the literature. However, the mechanisms involved to explain the effects of polyphenols are not elucidated yet. Therefore, the aim of this study was to investigate the mechanisms by which polyphenols from red wine (Provinols^TM) modulated contraction in human veins. We took advantage of a human model previously reported as new tool for pharmacological research using tissue-engineered techniques allowing the production of vascular media based exclusively on human smooth muscle cells. Thus, human tissue-engineered vascular media were produced with cells originating from umbilical cord vein. TEVM was treated either with vehicle or Provinols^TM. Results showed that treatment of TEVM by Provinols^TM significantly potentiated the contractile responses induced by histamine and bradykinin. The potentiating effect of Provinols^TM was not associated with an enhancement of histamine-induced increase in cytosolic calcium. It rather implied the presence of a Ca²⁺-independent signalling pathway. Pharmacological studies indicated that Provinols actions took place at the level of phospholipase A_2- Rho-kinase pathway and was associated with an enhancement of myosin light chain kinase activity. These results obtained using the human TEVM bring new insights to explain the regulation of venous contraction by polyphenols.