Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Ovarian AGE–RAGE/STAT3/AKT1‐related protein expression and phosphorylation. (A) Representative Western blot bands for AGE, RAGE, STAT3, AKT1, phospho‐STAT3 (Ser727), phospho‐AKT (Ser473), and GAPDH in the Control, Model, HLWDD‐L, HLWDD‐M, HLWDD‐H, and Metformin groups. (B) Relative densitometric analysis of AGE/GAPDH, RAGE/GAPDH, STAT3/GAPDH, AKT1/GAPDH, p‐STAT3/STAT3, and p‐AKT/AKT1. ** p < 0.01 vs. Control; ## p < 0.01 vs. Model. Densitometric comparisons were based on independent biological replicates and analyzed as described in Methods.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Analysis

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Molecular docking score matrix for the ten highest‐degree database‐derived candidate compounds and AKT1, STAT3, and RAGE. Values are predicted AutoDock Vina scores in kcal/mol; more negative values indicate more favorable predicted scores within this computational model. The matrix is presented for comparative hypothesis generation and does not establish high‐affinity binding, target engagement, or constituent‐level activity in vivo. Three representative complexes were subsequently evaluated by 100‐ns MD simulations (Figures ).

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Docking Assay, Derivative Assay, Binding Assay, Drug discovery, Activity Assay, In Vivo

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Ovarian AGE–RAGE/STAT3/AKT1‐related protein expression and phosphorylation. (A) Representative Western blot bands for AGE, RAGE, STAT3, AKT1, phospho‐STAT3 (Ser727), phospho‐AKT (Ser473), and GAPDH in the Control, Model, HLWDD‐L, HLWDD‐M, HLWDD‐H, and Metformin groups. (B) Relative densitometric analysis of AGE/GAPDH, RAGE/GAPDH, STAT3/GAPDH, AKT1/GAPDH, p‐STAT3/STAT3, and p‐AKT/AKT1. ** p < 0.01 vs. Control; ## p < 0.01 vs. Model. Densitometric comparisons were based on independent biological replicates and analyzed as described in Methods.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Analysis

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Working model integrating the computational and in vivo findings. Network pharmacology and docking prioritized AGE–RAGE/STAT3/AKT1‐related signaling as a candidate network; 100‐ns MD and MM/PBSA analyses supported the dynamic plausibility of three representative docked complexes. In vivo treatment was associated with reproductive, insulin‐related metabolic (including HOMA‐IR), inflammatory, total‐protein, and phosphorylation‐level changes. Dashed connectors denote association; direct target engagement and causal pathway mediation were not established.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: In Vivo, Docking Assay, Phospho-proteomics, Drug discovery

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: KPF ameliorates placental pathological changes and oxidative stress in GDM rats. (A) HE staining was performed to evaluate pathological alterations in placental tissues. Magnification: 100×; scale bar: 200 μm. (B) PAS staining was performed to assess glycogen distribution and deposition in placental tissues. Magnification: 100×; scale bar: 200 μm. (C, D) DHE staining was performed to evaluate ROS accumulation in placental tissues. Magnification: 200×; scale bar: 20 μm. (E–I) Western blot analysis was performed to determine the protein expression levels of AIM2, p-STAT3, GPX4, and SLC7A11 in placental tissues. N = 3, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: H&E Stain, Staining, Western Blot, Analysis, Expressing

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: KPF modulates trophoblast inflammatory responses and ferroptosis-related alterations through STAT3-associated signaling regulation. (A) Molecular docking analysis showing the potential interaction between KPF and STAT3 protein. (B, C) RT-qPCR analysis of STAT3 mRNA expression and transfection efficiency in HTR-8/SVneo cells. (D) CCK-8 assay was used to evaluate cell viability. (E, F) ELISA quantification of inflammatory cytokines IL-18 and TNF-α in cell supernatants. (G–L) Western blot analysis of AIM2, IL-1β, pro-IL-1β, GPX4, SLC7A11, cleaved caspase-1, and pro-caspase-1 protein expression in cells subjected to various treatments. (M–P) Western blot analysis of STAT3-associated changes in STAT3, GPX4, and SLC7A11 protein expression in cells subjected to various treatments. N = 3, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: Docking Assay, Analysis, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: STAT3-associated changes in STAT1/IRF1 signaling and AIM2 expression in HG-stimulated trophoblast cells. (A, B) RT-qPCR was performed to assess STAT3 knockdown efficiency and AIM2 mRNA expression changes in HTR-8/SVneo cells subjected to different treatments. (C–F) Western blot analysis was performed to evaluate the expression levels of p-STAT1, STAT1, IRF1, and AIM2 proteins under different experimental conditions. N = 3, not significant, ** P <0.01, *** P <0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot, Analysis