Dual endothelin receptor antagonists contrast the effects induced by endothelin-1 on cultured human microvascular endothelial cells

作者：Soldano S; Paolino S; Pizzorni C; Trombetta A C; Montagna P; Brizzolara R; Corallo C; Giordano N; Sulli A; Cutolo M

来源：Clinical and Experimental Rheumatology, 2017, 35(3): 484-493.

• endothelial cells
• endothelin-1
• TGF beta 1
• endothelial-to-mesenchymal transition
• dual endothelin receptor antagonists
• fibrotic process

Objective
To evaluate the ability of dual endothelin (ET) receptor antagonists (ETA/ETB - ETA/BRAs) to contrast the ET-1-induced effects on cultured human microvascular endothelial cells (HMVECs).
Methods
Some cultured HMVECs were untreated, or treated with ET-1 (100nM) or transforming growth factor beta 1 (TGF beta 1, 10 ng/mL) alone for 6 days, in order to induce the endothelial-to-mesenchymal transition (EndoMT). Other cultured HMVECs were pre-treated for 1hr with ETA/BRAs bosentan (10 mu M) or macitentan (1 mu M, 10 mu M) before the stimulation with ET-1 for 6 days. At the end of treatments, a mechanical injury was induced to cultured HMVECs (by scratching the cell monolayer with a sterile tip), and then the cell ability to re-fill the damaged area was determined after 24hrs. EndoMT phenotype markers and monocyte chemoattractant protein-1 (MCP-1) were evaluated by qRT-PCR and Western blotting. Statistical analysis was performed using Mann-Whitney-U non-parametric test.
Results
Both ET-1 and TGF beta 1 induced EndoMT and the MCP-1 over-expression in cultured HMVECs, as well as reduced the process of endothelial cell damage repair. Pre-treatment with ETA/BRAs let cultured HMVECs to significantly restore the in vitro damage of the cell monolayer and antagonised the EndoMT process as well as the MCP-1 over-expression (range p<0.05 - p<0.001). Conversely, untreated or TGF beta 1-treated HMVECs were found unaffected by the ETA/BRAs treatments.
Conclusion
The treatment with dual ETA/BRAs seems to partially restore the altered cell function induced by ET-1 in cultured endothelial cells, and might justify their therapeutic efficiency in clinical conditions characterised by increased concentrations of ET-1.

• 出版日期2017-6