lep  (Bioss)

Journal: Frontiers in Nutrition

Article Title: Astragaloside IV modulates oxidative stress and osteoimmune–Wnt signaling in ovariectomized rats: an integrated study of RNA sequencing, molecular docking, and experimental validation

doi: 10.3389/fnut.2026.1785452

Figure Lengend Snippet: Molecular docking models of Astragaloside IV (AS-IV) binding to the core targets. Representative binding poses of AS-IV (shown in stick representation) within the predicted binding pockets of the key target proteins. The calculated binding free energy (kcal/mol) for each complex is indicated. (A) Il22ra2 (−10.2 kcal/mol), (B) Ptgs2 (−9.5 kcal/mol), (C) Wnt1 (−9.3 kcal/mol), (D) Igfbp2 (−8.4 kcal/mol), (E) GZMB (−8.2 kcal/mol), (F) Lep (−7.8 kcal/mol), (G) Gfap (−7.8 kcal/mol), (H) Wnt10a (−7.7 kcal/mol). These results demonstrate stable binding interactions between the therapeutic compound AS-IV and all eight core targets identified from the PPI network and WGCNA, with Il22ra2 and Ptgs2 showing the strongest binding affinity.

Article Snippet: Primary antibodies diluted in antibody diluent were applied and incubated at 4 °C overnight; the primary antibodies were as follows: PTGS2 (ABclonal, A3560, rabbit, 1:200), IGFBP2 (Proteintech, 66,644-1-IG, mouse, 1:200), WNT1 (ABclonal, A2475, rabbit, 1:200), LEP (bioss, bs-0409R, rabbit, 1:200), GFAP (Proteintech, 60,190-1-IG, mouse, 1:1000), β -catenin (huilanbio, ABB3523, rabbit, 1:200), and NF-κB p65 (Cell Signaling Technology, 8,242, rabbit, 1:200), with EDTA-based retrieval used for all targets.

Techniques: Binding Assay

Journal: Frontiers in Nutrition

Article Title: Astragaloside IV modulates oxidative stress and osteoimmune–Wnt signaling in ovariectomized rats: an integrated study of RNA sequencing, molecular docking, and experimental validation

doi: 10.3389/fnut.2026.1785452

Figure Lengend Snippet: Multiplex immunofluorescence imaging and semi-quantification of target proteins in femoral trabecular bone. Representative multiplex immunofluorescence images showing DAPI (nuclei), target protein staining, and merged overlays in the femoral trabecular region from Sham, OVX, and OVX-AS-IV-M treated rats. Panels show staining for (A) NF-κB p65, (B) Wnt1, (C) Gfap, (D) β-catenin, (E) Igfbp2, (F) Lep, and (G) Ptgs2. Scale bar, 50 μm. The corresponding semi-quantification of mean fluorescence intensity (MFI) for each marker is presented to the right of each image panel, normalized to the Sham group (set to 1). Data are presented as mean ± SD with individual data points overlaid. Statistical comparisons were performed using one-way ANOVA followed by Dunnett’s multiple-comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies diluted in antibody diluent were applied and incubated at 4 °C overnight; the primary antibodies were as follows: PTGS2 (ABclonal, A3560, rabbit, 1:200), IGFBP2 (Proteintech, 66,644-1-IG, mouse, 1:200), WNT1 (ABclonal, A2475, rabbit, 1:200), LEP (bioss, bs-0409R, rabbit, 1:200), GFAP (Proteintech, 60,190-1-IG, mouse, 1:1000), β -catenin (huilanbio, ABB3523, rabbit, 1:200), and NF-κB p65 (Cell Signaling Technology, 8,242, rabbit, 1:200), with EDTA-based retrieval used for all targets.

Techniques: Multiplex Assay, Immunofluorescence, Imaging, Staining, Fluorescence, Marker

Journal: Redox Biology

Article Title: Ergothioneine rescues obesity-induced testicular dysfunction via dual restoration of steroidogenesis and mitochondrial redox homeostasis

doi: 10.1016/j.redox.2026.104090

Figure Lengend Snippet: Chronic high-fat diet progressively impairs testicular architecture and sperm quality. (A) Longitudinal body weight changes in control and HFD-fed mice over a 14-week period (left), and representative gross morphology showcasing significant obesity in the HFD group (right). n = 6. (B) Oral glucose tolerance test (OGTT) profiles demonstrating impaired glucose homeostasis in HFD mice. (C) Adipose and organ weights, including epididymal fat, inguinal fat, liver, brown adipose tissue (BAT), and testes, normalized to body weight. (D-E) Quantitative analysis of sperm concentration and total motility assessed by computer-assisted sperm analysis (CASA). (F) Representative H&E-stained sections of testicular tissues from SNC (Short-term Normal Control), SHFD (Short-term HFD), LNC (Long-term Normal Control), and LHFD (Long-term HFD) groups. scale bar = 50 μm. (G-H) Representative immunohistochemical (IHC) staining, morphometric quantification of the Mean Tubular Biopsy Score (MTBS) and germinal epithelium (GE) thickness, reflecting the degree of seminiferous tubule degeneration. (I-J) Quantification of Transition Protein 1 (TNP1) and γH2AX in testicular sections. γH2AX serves as a marker for DNA double-strand breaks in meiotic spermatocytes. scale bar = 20 μm. (K–N) Cytotoxicity profiles of mouse testicular cell lines (GC-1 spg, GC-2 spd, TM3, and TM4) following exposure to varying concentrations of palmitate (PA) for 48 h, assessed via CCK-8 assay. (O) Representative fluorescence images of BODIPY™ 493/503 staining visualizing neutral lipid droplet (LD) accumulation in GC-1 spg, GC-2 spd, TM3, and TM4 cells under lipotoxic conditions. scale bar = 20 μm. Data are presented as mean ± SEM ( n ≥ 3 independent biological replicates). The normality of the data was verified using the Shapiro-Wilk test. Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 indicate significant differences compared to the control group.

Article Snippet: The mice used in the study were sourced from two different suppliers, corresponding to two distinct animal models: i) The high-fat diet (HFD)-induced obesity model mice were obtained from Shanghai Model Organisms Center, Inc. (Shanghai, China).

Techniques: Control, Concentration Assay, Staining, Immunohistochemical staining, Immunohistochemistry, Marker, CCK-8 Assay, Fluorescence

Journal: Redox Biology

Article Title: Ergothioneine rescues obesity-induced testicular dysfunction via dual restoration of steroidogenesis and mitochondrial redox homeostasis

doi: 10.1016/j.redox.2026.104090

Figure Lengend Snippet: Progressive depletion of ergothioneine and lipid remodeling defined in the HFD testes . (A) Multivariate partial least squares-discriminant analysis (PLS-DA) score plot illustrating a distinct separation of metabolic phenotypes between the HFD and Control groups at different time points. (B) Volcano plot identifying significantly altered metabolites based on the criteria of variable importance in projection(VIP) > 1.0 and P < 0.05. (C) Chemical class distribution of the identified differential metabolites, highlighting major lipid and organic acid remodeling. (D) Differential abundance of top-ranked testicular metabolites, showing significantly upregulated and downregulated molecules across experimental cohorts. (E) Summary of the total number of differential metabolites identified through pairwise comparisons (SHFD vs. SNC; LHFD vs. LNC; and LHFD vs. SHFD), demonstrating the temporal metabolic shift. (F-G) Venn diagrams depicting intersectional analysis of shared and unique (F) upregulated and (G) downregulated metabolites among the indicated longitudinal comparisons. (H) Longitudinal quantification of l -Ergothioneine (ET) levels in testicular tissues across the SNC, SHFD, LNC, and LHFD groups, showcasing its progressive depletion during HFD-induced obesity. Data are presented as mean ± SEM ( n = 6 biological replicates for metabolomics). The normality of the metabolic data was rigorously assessed using the Shapiro-Wilk test. For multiple group comparisons, one-way ANOVA followed by Tukey's post-hoc test was employed. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus the respective control group.

Article Snippet: The mice used in the study were sourced from two different suppliers, corresponding to two distinct animal models: i) The high-fat diet (HFD)-induced obesity model mice were obtained from Shanghai Model Organisms Center, Inc. (Shanghai, China).

Techniques: Control