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. 2024 Oct 11;7(1):101242. doi: 10.1016/j.jhepr.2024.101242

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© 2024 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Fig. 1 — Frequency of Tregs is associated with HFD-induced hepatic steatosis and insulin resistance.

(A) Representative flow plots of the proportion of hepatic CD25⁺Foxp3⁺ Tregs gated on CD3⁺CD4⁺ T cells from mice fed a NCD or HFD (—four to five/group). (B) Number of hepatic CD4⁺CD25⁺Foxp3⁺ Tregs of mice fed a NCD or HFD for 1, 2, 4, and 6 weeks (five/group). (C, D) Foxp3 mRNA expression of liver tissues from (C) the NCD or HFD group (five/group), and (D) patients with or without hepatic steatosis (six/group). (E) Schematic of the construction of Treg-depleted mice (injected intraperitoneally with anti-CD25 antibodies) and Treg-expand mice (injected intraperitoneally with ldIL-2): representative dot plots of CD25⁺Foxp3⁺ Tregs gated on CD3⁺CD4⁺ T cells in the liver and a graph of the proportion of hepatic CD4⁺CD25⁺Foxp3⁺ Tregs (five/group). Comparison of (F) body weight (five/group), (G) GTT and ITT (five/group), (H) insulin-stimulated Akt and pAkt expression in the liver (three/group), (I) serum TC and total TG levels (five/group), and liver TC and TG levels (five/group) in Treg-depleted vs. Treg-expanded mice. (J) Representative images of H&E and Oil red O staining of liver sections (scale bars: 50 μm and 100 μm, respectively). (K) Representative images of Oil red O staining (scale bar: 100 μm) and TG content of primary hepatocytes co-cultured with Tregs or not (three/group). (L) Immunoblot analysis of insulin-stimulated Akt and pAkt in primary hepatocytes co-cultured with Tregs or not (three/group). Values represent means ± SD. Statistical analysis was performed by 2-tailed unpaired Student t test (B–D) or one-way ANOVA and Tukey's test (F–I,K,L): ∗p <0.05, ∗∗p <0.01, ∗∗∗p <0.001. Abbreviations: FFA, free fatty acid; GTT, glucose tolerance test; HFD, high-fat diet; ITT, insulin tolerance test; ldIL-2, low-dose IL-2; NCD, normal chow diet; pAkt, phosphorylated Akt; TC, total cholesterol; TG, triglyceride; Treg, regulatory T cell.

Fig. 1 — Frequency of Tregs is associated with HFD-induced hepatic steatosis and insulin resistance.

(A) Representative flow plots of the proportion of hepatic CD25⁺Foxp3⁺ Tregs gated on CD3⁺CD4⁺ T cells from mice fed a NCD or HFD (—four to five/group). (B) Number of hepatic CD4⁺CD25⁺Foxp3⁺ Tregs of mice fed a NCD or HFD for 1, 2, 4, and 6 weeks (five/group). (C, D) Foxp3 mRNA expression of liver tissues from (C) the NCD or HFD group (five/group), and (D) patients with or without hepatic steatosis (six/group). (E) Schematic of the construction of Treg-depleted mice (injected intraperitoneally with anti-CD25 antibodies) and Treg-expand mice (injected intraperitoneally with ldIL-2): representative dot plots of CD25⁺Foxp3⁺ Tregs gated on CD3⁺CD4⁺ T cells in the liver and a graph of the proportion of hepatic CD4⁺CD25⁺Foxp3⁺ Tregs (five/group). Comparison of (F) body weight (five/group), (G) GTT and ITT (five/group), (H) insulin-stimulated Akt and pAkt expression in the liver (three/group), (I) serum TC and total TG levels (five/group), and liver TC and TG levels (five/group) in Treg-depleted vs. Treg-expanded mice. (J) Representative images of H&E and Oil red O staining of liver sections (scale bars: 50 μm and 100 μm, respectively). (K) Representative images of Oil red O staining (scale bar: 100 μm) and TG content of primary hepatocytes co-cultured with Tregs or not (three/group). (L) Immunoblot analysis of insulin-stimulated Akt and pAkt in primary hepatocytes co-cultured with Tregs or not (three/group). Values represent means ± SD. Statistical analysis was performed by 2-tailed unpaired Student t test (B–D) or one-way ANOVA and Tukey's test (F–I,K,L): ∗p <0.05, ∗∗p <0.01, ∗∗∗p <0.001. Abbreviations: FFA, free fatty acid; GTT, glucose tolerance test; HFD, high-fat diet; ITT, insulin tolerance test; ldIL-2, low-dose IL-2; NCD, normal chow diet; pAkt, phosphorylated Akt; TC, total cholesterol; TG, triglyceride; Treg, regulatory T cell.