Journal: Redox Biology

Article Title: Development of SOCS1 mimetics as novel approach to harmonize inflammation, oxidative stress, and fibrogenesis in metabolic dysfunction-associated steatotic liver disease

doi: 10.1016/j.redox.2025.103670

Figure Lengend Snippet: S1 and liPS attenuate JAK/STAT and gene expression in livers of MCD diet-fed mice. (A) Representative immunoblots of P-STAT1/3, STAT1/3 and β-actin in liver samples from mice on MCD diet treated with vehicle (Ctrl) or peptides (S1 and liPS; 1.2 nmol/g), and normal diet (ND) as reference group. Quantitative analysis of P-STAT/STAT ratios expressed as fold increases versus ND group. (B) Representative images of P-STAT1/3 immunohistochemistry and quantification of positive area. Effect of peptides on MCD-induced mRNA levels of chemokines (C) , cytokines (D) , redox and ER-stress molecules (E) , and lipid transport genes (F) . The qPCR values were normalized by 18S. Results expressed as fold increases versus ND group are represented as individual data points and mean ± SD of the total number of animals per group. # P < 0.05 vs ND; ∗P < 0.05 vs Ctrl.

Article Snippet: Protein lysates (30–35 μg) from homogenized livers and cells were electrophoresed and immunoblotted for P-STAT1 (Thermo Fisher Scientific Cat# 33–3400, RRID: AB_2533113 ), STAT1 (Cell Signaling Technology Cat# 9172, RRID: AB_2198300 ), P-STAT3 (Cell Signaling) and STAT3 (Cell Signaling Technology Cat# 9139, RRID: AB_331757 ), with β-actin (Sigma-Aldrich Cat# A1978, RRID: AB_476692 ) and α-tubulin (Sigma-Aldrich Cat# T9026, RRID: AB_477593 ) as loading controls.

Techniques: Gene Expression, Western Blot, Immunohistochemistry

Journal: Redox Biology

Article Title: Development of SOCS1 mimetics as novel approach to harmonize inflammation, oxidative stress, and fibrogenesis in metabolic dysfunction-associated steatotic liver disease

doi: 10.1016/j.redox.2025.103670

Figure Lengend Snippet: Peptides reduce liver inflammation, oxidative stress, and fibrogenesis induced by MCD diet. (A) Representative images of F4/80, CD3, 8OHdG, and P-STAT1 immunoperoxidase, and Sirius red staining in liver sections from MCD diet-fed mice untreated (Control) and peptide-treated (S1, liPS, IC, and ICNal). (B) Quantification of positive stained area. Expression levels of inflammatory (C) , redox balance (D) and profibrotic (E) genes. The qPCR values normalized by 18S are expressed as fold increases versus normal diet (ND). Values are represented as individual data points and/or mean ± SD of the total number of animals per group. # P < 0.05 vs ND; ∗P < 0.05 vs Control (Co).

Article Snippet: Protein lysates (30–35 μg) from homogenized livers and cells were electrophoresed and immunoblotted for P-STAT1 (Thermo Fisher Scientific Cat# 33–3400, RRID: AB_2533113 ), STAT1 (Cell Signaling Technology Cat# 9172, RRID: AB_2198300 ), P-STAT3 (Cell Signaling) and STAT3 (Cell Signaling Technology Cat# 9139, RRID: AB_331757 ), with β-actin (Sigma-Aldrich Cat# A1978, RRID: AB_476692 ) and α-tubulin (Sigma-Aldrich Cat# T9026, RRID: AB_477593 ) as loading controls.

Techniques: Staining, Control, Expressing

Journal: Redox Biology

Article Title: Development of SOCS1 mimetics as novel approach to harmonize inflammation, oxidative stress, and fibrogenesis in metabolic dysfunction-associated steatotic liver disease

doi: 10.1016/j.redox.2025.103670

Figure Lengend Snippet: Hepatoprotective effect of peptides in a mouse model of HFD-induced MASLD/MASH. (A) Experimental design for testing peptides in HFD-fed ApoE −/− mice, with representative liver images. (B – C) Glucose tolerance test at 11 weeks of HFD in control (Ctrl), treated (S1, liPS, IC and ICNal) mice, and age-matched ND-fed mice. Blood glucose levels over time (B) and area under the curve (AUC) (C) were measured. (D) Representative images of hematoxylin/eosin, ORO, F4/80, 8OHdG, 4HNE, P-STAT1, and Sirius red staining in liver sections. (E) MASLD/NAFLD activity score. Quantification of lipid content (F) , F4/80+ macrophages (G) , oxidative stress markers (H) , P-STAT1 (I) , and collagen content (J) in liver samples. Values are represented as individual data points and/or mean ± SD of the total number of animals per group. # P < 0.05 vs ND; ∗P < 0.05 vs Ctrl.

Article Snippet: Protein lysates (30–35 μg) from homogenized livers and cells were electrophoresed and immunoblotted for P-STAT1 (Thermo Fisher Scientific Cat# 33–3400, RRID: AB_2533113 ), STAT1 (Cell Signaling Technology Cat# 9172, RRID: AB_2198300 ), P-STAT3 (Cell Signaling) and STAT3 (Cell Signaling Technology Cat# 9139, RRID: AB_331757 ), with β-actin (Sigma-Aldrich Cat# A1978, RRID: AB_476692 ) and α-tubulin (Sigma-Aldrich Cat# T9026, RRID: AB_477593 ) as loading controls.

Techniques: Control, Staining, Activity Assay

Journal: Redox Biology

Article Title: Development of SOCS1 mimetics as novel approach to harmonize inflammation, oxidative stress, and fibrogenesis in metabolic dysfunction-associated steatotic liver disease

doi: 10.1016/j.redox.2025.103670

Figure Lengend Snippet: Effects of linear and cyclic peptides in vitro. (A) Cell viability MTT assay in hepatocytes treated with 0.4 mM palmitic acid (PA) with/without peptides (S1, 59 μM; liPS, IC and ICNal, 25 μM). Medium with 10 % FBS was used as positive control. (B) Cell-based ELISA assay for STAT1 activation in hepatocytes stimulated with PA for 6 h in the presence of increasing concentrations of peptides. Values expressed as ratio P-STAT1/STAT1 versus PA alone. (C) NOX-dependent measurement of O2•− in hepatocytes after 16 h of stimulation, assessed by chemiluminescence assay. BSA and apocynin were used as controls. Lucigenin relative units are expressed as fold increases versus basal conditions. (D – F) Expression levels of redox, cell stress, lipid metabolism, and inflammatory genes in hepatocytes pretreated with peptides (S1, 59 μM; liPS, 25 μM; IC/ICNal, 12 and 25 μM) before stimulation with PA for 6 h (D) and 24 h (E and F) . (G) ELISA analysis of cytokine concentrations in hepatocyte conditioned media after 24 h of stimulation. Gene expression of M1 (H) and M2 (I) markers in macrophages incubated for 24 h with conditioned media from hepatocytes stimulated with PA with/without peptides. The qPCR values normalized by 18S are expressed as fold increases versus basal conditions. Results are represented as individual data points and/or mean ± SD of n = 4–6 independent experiments. # P < 0.05 vs basal; ∗P < 0.05 vs PA.

Article Snippet: Protein lysates (30–35 μg) from homogenized livers and cells were electrophoresed and immunoblotted for P-STAT1 (Thermo Fisher Scientific Cat# 33–3400, RRID: AB_2533113 ), STAT1 (Cell Signaling Technology Cat# 9172, RRID: AB_2198300 ), P-STAT3 (Cell Signaling) and STAT3 (Cell Signaling Technology Cat# 9139, RRID: AB_331757 ), with β-actin (Sigma-Aldrich Cat# A1978, RRID: AB_476692 ) and α-tubulin (Sigma-Aldrich Cat# T9026, RRID: AB_477593 ) as loading controls.

Techniques: In Vitro, MTT Assay, Positive Control, In-Cell ELISA, Activation Assay, Chemiluminescence Immunoassay, Expressing, Enzyme-linked Immunosorbent Assay, Gene Expression, Incubation