Characterization of connexin and pannexin expression in an in vivo and in vitro model of cholestasis

Abstract

Cholestasis is a liver disease, where the bile flow gets reduced or blocked and is characterized by an accumulation of bile acids. Cholestasis can be induced in different ways. In this thesis, an in vivo model as well as an in vitro model will be described. The in vivo model was generated after bile duct ligation in mice. The in vitro model was generated by adding the cholestatic drug, Atazanavir simultaneously with a mixture of bile acids to HepaRG cells.The aim of this study was to analyse the expression of Cx26, Cx32, Cx43 and Panx1 in cholestasis. An altered expression of these proteins in cholestasis could lead to the discovery of new biomarkers in the diagnosis of cholestasis. The expression of Cx26, Cx32, Cx43 and Panx1 was analysed in an in vivo as well as in an in vitro model of cholestasis. The expression on translational level was analysed using western blot. The subcellular localization of the proteins was analysed using IHC. Finally, the expression on transcriptional level was analysed using RT-qPCR. In the in vitro model, only Cx32 expression was analysed, using western blot. On translational level, an upregulation of Cx43 and Panx1 and a downregulation of Cx32 was observed in the cholestatic samples, generated by BDL. The upregulation of Cx43 and Panx1 was also observed on transcriptional level. The results of IHC are in line with the results of the western blot for Cx43 and Cx32. Finally, the expression of Cx32 was analysed in an in vitro model, but no significant results were generated. It can be concluded that during cholestasis, an upregulation of Cx43 and Panx1 takes place. In this way, Cx43 and Panx1 could be used as new biomarkers for the diagnosis of cholestasis. However, further research is required, especially for in the in vitro model.

Date of Award	2019
Original language	English
Supervisor	Axelle Cooreman (Advisor) & Mathieu Vinken (Promotor)