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The suitability of PPARγ and cytoglobin as markers for the reversion of the hepatic stellate cell in precision-cut liver slices.

Evering, Hester (2019) The suitability of PPARγ and cytoglobin as markers for the reversion of the hepatic stellate cell in precision-cut liver slices. Bachelor's Project, Pharmacy.

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Abstract

Background: In a healthy liver, hepatic stellate cells are present in the liver in a quiescent state, in which they store retinoids. During liver injury the hepatic stellate cell can be activated to an extracellular matrix proteins-producing cell (myofibroblast), under the influence of profibrotic growth factors, such as TGFβ. If the liver injury is solved, either drug-induced or by removal of the cause, the hepatic stellate cell can revert to a quiescent-like state. Peroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear receptor and its gene expression decreases during activation and increases during reversion. Cytoglobin is an O2-transporter and ROS-scavenger and its gene expression decreases during activation in cell lines and increases during reversion. The aim of this study was to evaluate whether PPARγ and cytoglobin are possible markers for the reversion of the hepatic stellate cell in precision-cut liver slices (PCLS). Materials and Methods: PCLS were made from the livers of three adult male C57BL/6 mice. They are incubated in Williams medium E (with an addition of glucose and gentamycin) and with or without addition of TGFβ (5ng/mL) and/or Galunisertib (10 μM) for 24 hours or 48 hours. The slices were analysed by measuring the ATP and protein concentration for the viability and by measuring the gene expression of collagen 1A1, PPARγ and cytoglobin with Real-time PCR. Results and Discussion: The gene expression of collagen 1A1 increased after induction of the fibrosis with TGFβ and decreased during resolution, drug-induced or by removal of TGFβ. The expression of PPARγ increased after 24 hours stimulation with TGFβ. After 48 hours of induction of fibrosis with TGFβ or resolution of fibrosis with Galunisertib the gene expression of PPARγ was not altered. The difference between the results in PCLS and cell lines could be caused by the other cell types, that are present in PCLS, for example the hepatocytes. The expression of cytoglobin increased after stimulation with TGFβ and decreased after induced resolution of the fibrosis with Galunisertib, which could be caused by the role of cytoglobin as O2-transporter and ROS-scavenger. Conclusion: Addition of an antifibrotic drug (Galunisertib) or removal of the profibrotic stimulus leads to resolution of the fibrosis in PCLS as shown by the changes in collagen 1A1 levels. However, PPARγ and cytoglobin are not suitable markers to analyse the reversion of hepatic stellate cells in PCLS. Therefore, in future studies other markers for the reverted hepatic stellate cell should be tested in this slice system.

Item Type: Thesis (Bachelor's Project)
Supervisor:
Supervisor nameSupervisor E mail
Beljaars, E.E.Beljaars@rug.nl
Degree programme: Pharmacy
Thesis type: Bachelor's Project
Language: French
Date Deposited: 03 Jul 2019
Last Modified: 04 Jul 2019 09:33
URI: http://fse.studenttheses.ub.rug.nl/id/eprint/19837

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