2.8. Cholesterol Synthesis The reaction steps from mevalonate to farnesyl-diphosphate are down-regulated with time, and further down-regulated by TGFβ treatment, see Figure 5A. The 12 reaction steps from
Farnesyl-diphosphate show a similar pattern (see Figure 5A and B), which is a considerable down-regulation with time and further down-regulation by TGFβ treatment, with a single exception—lathosterol oxidase (Sc5d) is up-regulated by TGFβ. However, this multi-specific enzyme (alternative substrates are for example δ7-avenasterol and episterol) is also involved in other BYL719 supplier relevant functions. Figure 5 Regulation of genes involved in cholesterol synthesis. Conclusively, cholesterol synthesis can Inhibitors,research,lifescience,medical be predicted as down-regulated Inhibitors,research,lifescience,medical in the control experiment and even more down-regulated
in TGFβ treated hepatocytes. This is not surprising as cholesterol synthesis (for bile acids and lipoprotein particles export) is a typical liver function. 2.9. Glucose Release from Glycogen The reactions involved in the hepatic release of glucose from glycogen storage can be grouped in two—split of activated glucose Inhibitors,research,lifescience,medical from the glycogen polysaccharide structure and dephosphorylation and export of glucose. As can be seen from Figure 6A, there is only a slight down-regulation of the first group of genes (Pygl, Pgm2) while the second group (Slc37a4, G6pc, Slc2a2) is sharply down-regulated with time, in particular in TGFβ treated hepatocytes. This is agreement with macroscopic observations— degradation of the cell’s glycogen storage is a universal function of human cells while the actual export of glucose is
specific for Inhibitors,research,lifescience,medical hepatocytes. In particular, dephosphorylation of glucose-6-phosphate Inhibitors,research,lifescience,medical (G6pc) (a reaction only needed for glucose export) switches from a clearly on to an off status. Figure 6 Regulation of genes involved in glucose release from glycogen. Glucose-6-phosphatase shows an early down-regulation in the control experiment (from 1 h to 6 h), while in TGFβ treated sample, down-regulation occurs later, i.e., in the interval between 6 h and 24 h (Figure 6B). From this result, it can be hypothesized that loss of Thiamine-diphosphate kinase glucose export capability is delayed in the TGFβ treated hepatocytes. 2.10. Supply of β-hydroxybutyrate Genes involved in the β-hydroxybutyrate synthesis pathway of show an inconclusive regulation when comparing 1 h to 24 h. While mitochondrial HMG-CoA synthase (Hmgcs2) is down-regulated in the control experiments and up-regulated upon TGFβ treatment, both types of 3-hydroxybutyrate dehydrogenase (Bdh1/Bdh2) are up-regulated in the control experiment and unchanged in TGFβ treated hepatocytes. Intriguingly, the two genes Acat1 and Hmgcs2 show a rare pattern in TGFβ treated sample, that is they are up-regulated at the 6 h time point and then down-regulated again (see Figure 7B).