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. 2020 Sep 30;319(6):G669–G684. doi: 10.1152/ajpgi.00175.2020

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Fig. 4. — Fibroblast growth factor 15 (Fgf15) deficiency is associated with accumulation of esterified cholesterol and lipotoxicity in mice post-vertical sleeve gastrectomy (VSG). A: hepatic triglyceride (TG) content 49 days postsurgery. Liver triglyceride levels were much lower in both VSG groups compared with their respective Sham controls; n: wild-type (WT) Sham = 6, WT VSG = 6, knockout (KO) Sham = 5, KO VSG = 7. **P < 0.01, one-way ANOVA. B: liver weight/body weight ratio 49 days postsurgery. Liver weight/body weight ratio was greater in KO VSG mice compared with all other groups; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, **P < 0.01, one-way ANOVA. C: hepatic free cholesterol content 49 days postsurgery. Liver free cholesterol levels were elevated in both VSG groups; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, one-way ANOVA. D: hepatic esterified cholesterol content 49 days postsurgery. Liver esterified cholesterol levels were elevated in KO VSG group; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, one-way ANOVA, t test. E: Hmg-CoA reductase (3-hydroxy-3-methyl-glutaryl-CoA reductase), Abcg5, and Abcg8 gene expression at day 49 postsurgery. mRNA levels of the genes coding for the cholesterol synthesis (Hmg-CoA reductase), cholesterol efflux pump ATP-binding cassette, subfamily G (WHITE), member 5 (sterolin 1, Abcg5), cholesterol efflux pump ATP-binding cassette, subfamily G (WHITE), member 8 (sterolin 2, Abcg8) were measured by RT-PCR and expressed in relative expression units. Cholesterol synthesis rate-limiting gene was not downregulated in KO VSG group. Cholesterol export genes were suppressed in KO VSG group; n: WT Sham = 4, WT VSG = 4, KO Sham = 4, KO VSG = 6. *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA, t test). F: Cpt1a, CD36, and Scd1 gene expression at day 49 postsurgery. mRNA levels of the genes coding lipid transport carnitine palmitoyltransferase-1A (Cpt1a), lipid synthesis (CD36), and lipid synthesis stearoyl-CoA desaturase (Scd1) were measured by RT-PCR and expressed in relative expression units. These genes were mostly suppressed in WT VSG group (only trend in KO VSG) relative to Sham group; n: WT Sham = 4, WT VSG = 5, KO Sham = 4, KO VSG = 6. *P < 0.05, ***P < 0.0001, one-way ANOVA, t test. G: hepatic fatty acid oxidation gene expression at day 49 postsurgery. mRNA levels of genes coding for fatty acid oxidation were measured by Biomark HD System high-throughput PCR and expressed in relative expression units. Acc1 (acetyl-CoA carboxylase-1), Acc2, and Fasn (fatty acid synthase) genes were significantly overexpressed only in KO VSG mice; n: WT Sham = 4, WT VSG = 5, KO Sham = 4, KO VSG = 6; *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA, t test; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7; *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA. H: heat map of significantly changed lipid compounds obtained from WT VSG vs. KO VSG mouse livers. Phosphatidylcholine (PC) was decreased in KO VSG comparing to WT VSG, while phosphatidylinositol (PI), diacylglycerol (DAG), phosphatidylglycerol (PG), and sphingomyelin (SM) were increased in KO VSG mice; n: WT VSG = 5, KO VSG = 5. I: volcano plot of 418 identified lipid compounds; 24 lipid compound ions were significantly different between WT VSG and KO VSG groups with fold change >2 and P < 0.05; n: WT VSG = 5, KO VSG = 5. J: principal component analysis (PCA) of mouse liver separates; 418 identified lipid compounds were analyzed in electrospray positive and negative modes from liver tissue of mice collected at 49 days postsurgery; n: WT VSG = 5, KO VSG = 5.

Fig. 4. — Fibroblast growth factor 15 (Fgf15) deficiency is associated with accumulation of esterified cholesterol and lipotoxicity in mice post-vertical sleeve gastrectomy (VSG). A: hepatic triglyceride (TG) content 49 days postsurgery. Liver triglyceride levels were much lower in both VSG groups compared with their respective Sham controls; n: wild-type (WT) Sham = 6, WT VSG = 6, knockout (KO) Sham = 5, KO VSG = 7. **P < 0.01, one-way ANOVA. B: liver weight/body weight ratio 49 days postsurgery. Liver weight/body weight ratio was greater in KO VSG mice compared with all other groups; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, **P < 0.01, one-way ANOVA. C: hepatic free cholesterol content 49 days postsurgery. Liver free cholesterol levels were elevated in both VSG groups; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, one-way ANOVA. D: hepatic esterified cholesterol content 49 days postsurgery. Liver esterified cholesterol levels were elevated in KO VSG group; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7. *P < 0.05, one-way ANOVA, t test. E: Hmg-CoA reductase (3-hydroxy-3-methyl-glutaryl-CoA reductase), Abcg5, and Abcg8 gene expression at day 49 postsurgery. mRNA levels of the genes coding for the cholesterol synthesis (Hmg-CoA reductase), cholesterol efflux pump ATP-binding cassette, subfamily G (WHITE), member 5 (sterolin 1, Abcg5), cholesterol efflux pump ATP-binding cassette, subfamily G (WHITE), member 8 (sterolin 2, Abcg8) were measured by RT-PCR and expressed in relative expression units. Cholesterol synthesis rate-limiting gene was not downregulated in KO VSG group. Cholesterol export genes were suppressed in KO VSG group; n: WT Sham = 4, WT VSG = 4, KO Sham = 4, KO VSG = 6. *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA, t test). F: Cpt1a, CD36, and Scd1 gene expression at day 49 postsurgery. mRNA levels of the genes coding lipid transport carnitine palmitoyltransferase-1A (Cpt1a), lipid synthesis (CD36), and lipid synthesis stearoyl-CoA desaturase (Scd1) were measured by RT-PCR and expressed in relative expression units. These genes were mostly suppressed in WT VSG group (only trend in KO VSG) relative to Sham group; n: WT Sham = 4, WT VSG = 5, KO Sham = 4, KO VSG = 6. *P < 0.05, ***P < 0.0001, one-way ANOVA, t test. G: hepatic fatty acid oxidation gene expression at day 49 postsurgery. mRNA levels of genes coding for fatty acid oxidation were measured by Biomark HD System high-throughput PCR and expressed in relative expression units. Acc1 (acetyl-CoA carboxylase-1), Acc2, and Fasn (fatty acid synthase) genes were significantly overexpressed only in KO VSG mice; n: WT Sham = 4, WT VSG = 5, KO Sham = 4, KO VSG = 6; *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA, t test; n: WT Sham = 6, WT VSG = 6, KO Sham = 5, KO VSG = 7; *P < 0.05, **P < 0.01, ****P < 0.0001, one-way ANOVA. H: heat map of significantly changed lipid compounds obtained from WT VSG vs. KO VSG mouse livers. Phosphatidylcholine (PC) was decreased in KO VSG comparing to WT VSG, while phosphatidylinositol (PI), diacylglycerol (DAG), phosphatidylglycerol (PG), and sphingomyelin (SM) were increased in KO VSG mice; n: WT VSG = 5, KO VSG = 5. I: volcano plot of 418 identified lipid compounds; 24 lipid compound ions were significantly different between WT VSG and KO VSG groups with fold change >2 and P < 0.05; n: WT VSG = 5, KO VSG = 5. J: principal component analysis (PCA) of mouse liver separates; 418 identified lipid compounds were analyzed in electrospray positive and negative modes from liver tissue of mice collected at 49 days postsurgery; n: WT VSG = 5, KO VSG = 5.