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Huh7.5 Cells Grown in Human Serum as a Model to Study the Effects of Hepatitis C Virus Infection on Lipid Catabolism

  • Author / Creator
    Tat, Wilson
  • Our goal is to study the additional elements of the mechanism of steatosis underlying hepatitis C virus (HCV) infection. Human hepatoma Huh7.5 cells cultured in human serum (HS) were used as a model to study the development of steatosis. Huh7.5 cells grown in HS media were chosen to study the effects of HCV infection on lipid catabolism due to the increased expression of transcription factors associated with lipid catabolism compared to cells grown in fetal bovine serum (FBS) media. Here we show that cells grown in HS media have increased degradation of fatty acids through β-oxidation compared to cells grown in FBS media. This result was reflected by the increase in protein levels of carnitine palmitoyltransferase 1 (CPT-1), the rate limiting step of β-oxidation. Additionally, there was significantly higher production of ketone bodies in cells grown in HS media relative to FBS media. These ketone bodies are generated from the ketogenesis pathway that is downstream of β-oxidation. Finally, when these cells were grown in HS media compared to FBS media, there were larger lipid droplets and significantly more triglyceride. These results show that Huh7.5 cells cultured in HS have increased lipid metabolism compared to cells grown in FBS media. During HCV infection, we showed that the rate of β-oxidation was significantly decreased. This reduction in β-oxidation was associated with the reduction of CPT-1 and reduction of ketone body production. The reduction in lipid catabolism observed during HCV infection was reflected with the increased lipid content in HCV infected cells compared to uninfected cells. These results indicate that HCV infection leads to lower β-oxidation, potentially contributing to lipid accumulation and steatosis during HCV infection.

  • Subjects / Keywords
  • Graduation date
    2018-06
  • Type of Item
    Thesis
  • Degree
    Master of Science
  • DOI
    https://doi.org/10.7939/R32R3PC1W
  • License
    Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.