Aims: The endothelial cells (EC) play a crucial role in the regulation of a broad range of many physiological and pathological processes. The aims of this work are to evaluate possible gender differences in EC obtained by male and female newborn umbilical cord in cell fate and in the ability of cell survival in starvation an inducer of autophagy. Methods: HUVECs (human umbilical vein endothelial cells) were isolated as previously described by Cheung L et al. and umbilical cord were provided by the department of Pharmacology, Gynecology, and Obstetrics by University of Sassari. The cell proliferation was evaluated with MTS assay kit and dimension and cell shape were evaluated with Labworks sofware. Moreover we determinate H2O2 LDH and PAI-1 concentration with commercial colorimetric assay kit respectively in cell supernatants and, eNOS, Beclin1, LC3, AKT, mTOR expression by western blotting on cell lysates according to the instructions provided by Cell Signalling Tecnology assay kit and nitrite quantification by Griess reaction on culture supernatants. The experiments were performed with the 5 passage. Results: Basically, no significant differences are observed in cell shape and dimension, LDH, PAI-1 and nitrite and nitrate production levels and espression of eNOS. But basal male cells have a higher expression of Beclin1, an autophagic marker, while the expression of AKT and its activity as mTOR expression are significantly higher in female than in male cells. After 36 hours of starvation, as expected both cell types are more autophagic, but male cells shows significantly higher expression of ratio LC3II/LC3I, an autophagic activity indicator. Finally, both cell types have lower AKT and mTOR espression and both cells show no AKT activity. Conclusion: When intracellular nutrients are limited as during starvation, eukaryotic cells can trigger a pathway of survival called autophagy. Here, it is evident that male and female endothelial cells have different responses.

Le Cellule endoteliali del cordone ombelicale hanno un destino cellulare che dipende dal sesso del feto / Addis, Roberta. - (2011 Feb 28).

Le Cellule endoteliali del cordone ombelicale hanno un destino cellulare che dipende dal sesso del feto

ADDIS, Roberta
2011-02-28

Abstract

Aims: The endothelial cells (EC) play a crucial role in the regulation of a broad range of many physiological and pathological processes. The aims of this work are to evaluate possible gender differences in EC obtained by male and female newborn umbilical cord in cell fate and in the ability of cell survival in starvation an inducer of autophagy. Methods: HUVECs (human umbilical vein endothelial cells) were isolated as previously described by Cheung L et al. and umbilical cord were provided by the department of Pharmacology, Gynecology, and Obstetrics by University of Sassari. The cell proliferation was evaluated with MTS assay kit and dimension and cell shape were evaluated with Labworks sofware. Moreover we determinate H2O2 LDH and PAI-1 concentration with commercial colorimetric assay kit respectively in cell supernatants and, eNOS, Beclin1, LC3, AKT, mTOR expression by western blotting on cell lysates according to the instructions provided by Cell Signalling Tecnology assay kit and nitrite quantification by Griess reaction on culture supernatants. The experiments were performed with the 5 passage. Results: Basically, no significant differences are observed in cell shape and dimension, LDH, PAI-1 and nitrite and nitrate production levels and espression of eNOS. But basal male cells have a higher expression of Beclin1, an autophagic marker, while the expression of AKT and its activity as mTOR expression are significantly higher in female than in male cells. After 36 hours of starvation, as expected both cell types are more autophagic, but male cells shows significantly higher expression of ratio LC3II/LC3I, an autophagic activity indicator. Finally, both cell types have lower AKT and mTOR espression and both cells show no AKT activity. Conclusion: When intracellular nutrients are limited as during starvation, eukaryotic cells can trigger a pathway of survival called autophagy. Here, it is evident that male and female endothelial cells have different responses.
Huvec; destino celluare; autofagia
Le Cellule endoteliali del cordone ombelicale hanno un destino cellulare che dipende dal sesso del feto / Addis, Roberta. - (2011 Feb 28).
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11388/251014
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