The Uremic Toxin Parathyroid Hormone Induces Endothe-Lial-mesenchymal Transition In Vascular Endothelial Cells And Its Molecular Mechanism

Objective: To investigate whether uremic toxins parathyroid hormone (PTH) couldinduce endothelial-mesenchymal transition (EndMT) in vascular endothelial cell and itsrelated mechanisms.Methods: As Human Aortic Endothelial Cells (HAEC) were cultured to logarithmicphase, PTH was added into the cultures with different doses (10-12,10-11,10-10,10-9,10-8mol/L) for48h; or with10-8mol/L PTH for12h,24h,36h and60h. Then the morphologicalchanges of HAEC cells were observed and photoed. The vascular endothelial cell markersVE-Cadherin and CD31, and the mesenchymal marker α-SMA were detected with real-timePCR and western blot. Furthermore, the expression of TGF-β1and ILK was investigated bywestern blot. TGF-β1signaling inhibitor (SB431542and Pirfenidone) and ILK inhibitorCpd22were used to investigate the potential mechanism of EndMT induced by PTH inHAECs.Results: The results of cell morphology suggested that HAECs have translated tomesenchymal-like cells induced by PTH. Real-time PCR and western blot results showedthat PTH reduced the expression of vascular endothelial cell marker CD31andVE-Cadherin. On the other hand, PTH was able to significantly increase the expression offiber cell marker α-SMA by a time-and dose-dependent manner. In addition, western blotanalysis showed that the expression of TGF-β1and ILK simultaneously increased upon thetreatment of PTH in HAECs. Furthermore, the TGF-β1signaling inhibitor (SB431542andPirfenidone) and ILK inhibitor (Cpd22) were able to reverse the EndMT induced by PTH inHAECs.Conclusion: PTH was able to induce the EndMT characteristics of HAEC cells, andTGF-β1/ILK signal pathway may play an important role in the EndMT induced by PTH inHAECs.