Heart failure (HF) is characterized by inflammatory conditions which cause endothelium dysfunctions and promote a worsening of the disease. We have previously shown that the serum of HF patients has a pro-apoptotic effect on human umbilical vein endothelial cells (HUVECs) which correlates with circulating levels of TNF-α. TNF-α dysregulates Notch signalling in HUVECs causing apoptosis. To establish whether the pro-apoptotic effect of HF serum in HUVECs could be due to dysregulation of Notch signalling, HUVECs were treated with serum of patients with mild and moderate/severe HF (New York Heart Association (NYHA) class I/II and III/IV respectively) or healthy controls and the effects on Notch signalling were evaluated. Western blot analyses showed reduced levels of Notch4 receptor in cells treated with serum from NYHA class I/II patients (3/12) and NYHA class III/IV (7/12) compared to control. Similarly, in serum-treated cells the mature form of Notch ligand Delta Like-4 (Dll4) was absent (2/12 class NYHA I/II and 7/12 class III/IV). Only with serum from class III/IV patients we found both reduced levels of Notch 4 and absence of Dll4 (4/12). Notch 1, 2 as well as Jagged1 ligand protein levels were unchanged by treatment. qRT-PCR showed increased Hey2 and reduced Hes1 and Hey1 mRNAs in treated cells compared to controls. These differences in mRNAs didn’t correlate with the severity of disease. In conclusion Notch signalling in HUVECs is modulated by inflammatory cytokines in serum of HF patients indicating that dysregulation of this pathway could be part of a mechanism by which these cytokines induce endothelial dysfunctions in HF patients.
Serum from heart failure patients modulates Notch signalling in human umbelical vein endothelial cells.
CALICETI, CRISTIANA;HRELIA, SILVANA;LEONCINI, EMANUELA;
2013
Abstract
Heart failure (HF) is characterized by inflammatory conditions which cause endothelium dysfunctions and promote a worsening of the disease. We have previously shown that the serum of HF patients has a pro-apoptotic effect on human umbilical vein endothelial cells (HUVECs) which correlates with circulating levels of TNF-α. TNF-α dysregulates Notch signalling in HUVECs causing apoptosis. To establish whether the pro-apoptotic effect of HF serum in HUVECs could be due to dysregulation of Notch signalling, HUVECs were treated with serum of patients with mild and moderate/severe HF (New York Heart Association (NYHA) class I/II and III/IV respectively) or healthy controls and the effects on Notch signalling were evaluated. Western blot analyses showed reduced levels of Notch4 receptor in cells treated with serum from NYHA class I/II patients (3/12) and NYHA class III/IV (7/12) compared to control. Similarly, in serum-treated cells the mature form of Notch ligand Delta Like-4 (Dll4) was absent (2/12 class NYHA I/II and 7/12 class III/IV). Only with serum from class III/IV patients we found both reduced levels of Notch 4 and absence of Dll4 (4/12). Notch 1, 2 as well as Jagged1 ligand protein levels were unchanged by treatment. qRT-PCR showed increased Hey2 and reduced Hes1 and Hey1 mRNAs in treated cells compared to controls. These differences in mRNAs didn’t correlate with the severity of disease. In conclusion Notch signalling in HUVECs is modulated by inflammatory cytokines in serum of HF patients indicating that dysregulation of this pathway could be part of a mechanism by which these cytokines induce endothelial dysfunctions in HF patients.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
