| Abstract: | BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease in Western society. Comparative gene expression studies are beginning to elucidate the molecular mechanisms underlying NAFLD progression. We have previously shown that high fat diets during early life can prime non-alcoholic steatohepatitis (NASH) in adulthood, through lipogenesis gene elevation. To generate accurate results in such studies, appropriate housekeeping genes (HKG) which are unaffected by disease processes, are used for data normalisation. However, there is little existing data to show the effects of NAFLD on HKG expression. AIMS: To identify the HKG in a mouse model of developmentally primed NAFLD and NASH, which maintains expression stability. METHODS: We determined the expression stability of six candidates HKG (GAPDH, YWHAZ, B2M, EIF4A2, ACTB and CYC1) in a mouse model of developmentally primed NAFLD in both the day and night, using geNORM qBasePlus software. RESULTS: HKG expression differed across dietary groups and time of day. In the majority of treatment groups and time points the most stable gene was YWHAZ. Following high fat diet interventions CYC1 became notably unstable. Overall the effect of NAFLD and NASH on HKG expression was to maintain stability of YWHAZ, but destabilise CYC1 and EIF4A2. CONCLUSIONS: Our data clearly shows that HKG expression is affected by NAFLD severity and time of day sampling, highlighting the importance of suitable HKG gene selection. For comparative gene expression studies investigating NAFLD we would recommend use of YWHAZ as a robust, stably expressed HKG. |
