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. 2022 Nov 16;9:1051157. doi: 10.3389/fnut.2022.1051157

FIGURE 3.

FIGURE 3

L-Ornithine L-Aspartate (LOLA) does not improve hepatic ammonia detoxification. (A) LOLA was administered in drinking water to HFD-fed FOZ mice (HFD + LOLA) for the last 8 weeks of the 12 weeks dietary experiment. Treated mice were compared with FOZ HFD given plain water (HFD). (B) mRNA levels of glutaminase (GLS) 1 and 2, of urea cycle enzymes (UCEs): carbamoyl phosphate synthetase (CPS1), ornithine transcarbamylase (OTC), Argininosuccinate synthetase (ASS1), argininosuccinate lyase (ASL), arginase (ARG1), and of glutamine synthetase (GS) in liver tissue from HFD and HFD + LOLA (n = 4–6); (C) Enzyme activity of OTC in liver tissue from HFD and HFD + LOLA (n = 4–6); (D) mRNA levels of GLS1 and GS in muscle tissue (gastrocnemius) from HFD and HFD + LOLA (n = 4–6); (E) mRNA levels of GLS1 in duodenum from HFD and HFD + LOLA (n = 4–6); (F) GS protein expression levels assessed by immunofluorescence in liver tissue from HFD and HFD + LOLA and (G) quantification of GS-positive tissue (in percentage) in liver sections (n = 4–6). (H) Plasma ammonia concentrations measured in systemic blood from HFD and HFD + LOLA (n = 4–6); (I) Representative hematoxylin-eosin (H&E) and Sirius red (SR) staining of liver sections from HFD and HFD + LOLA; (J) Non-alcoholic fatty liver disease activity score (NAS) (n = 4–6); (K) Liver-to-spleen density measured in vivo by micro-CT in HFD and HFD + LOLA after 12 weeks of diet (n = 4–6). All data are presented as mean ± SD, *p < 0.05, **p < 0.01. Statistical tests used: (C,E,G,J,K) Unpaired two-tailed t-test. (H) Repeated-measures two-way ANOVA and (B,D) two-way ANOVA followed by post hoc Bonferroni correction.