Franny Jongbloed

122 CHAPTER 5 Transcriptional responses after a methionine-, leucine- and tryptophan- free diet are highly overlapping To examine transcriptome responses amongst the different EAA-free diets and their controls, a principal component analysis (PCA) was made based on all probe sets in the microarray data (Figure 3A). 6 24 0 5000 10000 15000 * * ** ** ** * ** Time (hours) ALT (mmol/L) 6 24 0 20000 40000 60000 Control (n=6) Protein-free (n=6) Leucine-free (n=6) Methionine-free (n=6) Tryptophan-free (n=6) 3d 30% DR (n=8) * ** ** ** * ** * Time (hours) LDH (mmol/L) 0 20 40 60 80 100 120 Control (n=6) Protein-free (n=6) Tryptophan-free (n=6) 3d 30% DR (n=8) Leucine-free (n=6) Methionine-free (n=6) 6 24 * * * * * * Time (hours) neutrophils/ microscopic field A B C D 0 20 40 60 80 6 24 * * * * * * Time (hours) Necrosis (%) Figure 2. Essential amino acid free diets reduce liver injury. (A) Serum concentration of the liver-specific enzyme alanine aminotransferase (ALT), indicative of liver damage, were significantly lower levels at six and 24 hours after IRI in mice fed a protein-free or amino acid-free diet. *P<0.05, **P<0.01 versus the control group at the same time point. (B) Twenty-four hours after IRI, serum concentration of lactate dehydrogenase (LDH), indicative of tissue breakdown, was significant lower in mice on a protein-free or amino acid-free diet. *P<0.05, **P<0.01 versus the control group at the same time point. (C) Six hours after reperfusion, livers from mice fed leucine-free and tryptophan-free diets showed less hemorrhagic necrosis compared to the control group. Twenty- four hours after reperfusion the protein-, leucine- and methionine-free diets induced significantly less damage compared to the control group. *P<0.05, **P<0.01 versus the control group. (D) At 24 hours post-reperfusion, the protein-free diet had a lower number of neutrophils than the control group, while the 30% DR remained with a significantly higher neutrophil count. *P<0.05, versus the control group.