Journal: Frontiers in Pharmacology

Article Title: Huangkui capsule combined with finerenone attenuates diabetic nephropathy by regulating the JAK2/STAT3 signaling pathway based on network pharmacology, molecular docking, and experimental verification

doi: 10.3389/fphar.2025.1625286

Figure Lengend Snippet: Network pharmacology and molecular docking analysis of HKC and Finerenone in DN treatment. (A, B) Venndiagrams of drug-disease overlapping targets. HKC and Finerenone shared 137 common targets with DN. (C) Drug-component-target-disease network. Top three HKC component Degrees: Quercetin (101), Myricetin (101), Gossypetin (101). (D, E) PPI network and core targets. 11 key targets (JAK2, STAT3, AKT1, etc.) identified via CytoHubba. (F) Subnetwork of top-ranked targets from MCC algorithm analysis, highlighting high-confidence interactions. (G) Clustering analysis of PPI network using MCODE, showing densely connected target modules. (H) Heatmap of topological parameter scores (BC, CC, DC) for core targets. (I, J) GO/KEGG enrichment. JAK/STAT pathway was the most enriched. (K) Bubble plot of KEGG pathways ranked by significance (-log10 (p-value)). (L–N) GSE30122 analysis. JAK2/STAT3 expression elevated in DN renal tubules (logFC≥1.5, p<0.05). (O) Molecular docking binding energy. STAT3-Quercetin complex showed strongest binding affinity. (P–S) 3D and 2D interaction diagram of quercetin-JAK2/STAT3 and finerenone-JAK2/STAT3 binding pocket with hydrogen bond formation. (T) Root Mean Square Deviation (RMSD) trajectory of molecular dynamics simulations for protein-ligand complexes. (U) Root Mean Square Fluctuation (RMSF) analysis of amino acid residues in simulated complexes. (V) Radius of gyration (Rg) plot demonstrating structural stability during simulations. (W) Hydrogen bond occupancy analysis between core components and target proteins over 100ns simulation.

Article Snippet: HK-2 cells were induced with LPS (10 μg/mL) (MCE, United States) combined with high glucose (30 mmol/L) to establish inflammation and apoptosis models ( ) and then treated with quercetin (Que) and finerenone (Fine) (MCE, United States, drugs were dissolved in DMSO, concentration 0.1%).

Techniques: Expressing, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Huangkui capsule combined with finerenone attenuates diabetic nephropathy by regulating the JAK2/STAT3 signaling pathway based on network pharmacology, molecular docking, and experimental verification

doi: 10.3389/fphar.2025.1625286

Figure Lengend Snippet: Drug toxicity of quercetin and finerenone on H-K2 cells: (A,B) (n = 6):CCK-8 assay was used to detect the drug toxicity and cell survival rate of HK-2 cells treated with quercetin and finerenone for 24 and 48 h, respectively. Synergistic effect of quercetin and finerenone in HK-2 cells. (C) Dose–effect curve, IC50 (quercetin = 50.75 μM, finerenone = 75.48 μM). (D) Chou–Talalay analysis. Combination index (CI = 0.80; Cl > 1, antagonistic effect; Cl = 1, additive effect; Cl < 1, synergistic effect) confirmed synergy. (E) Equivalent dose curve; the combination of quercetin and finerenone exhibited significant synergistic effects at inhibition rates of 50%, 75%, and 90%. (F) Median effect curve.

Article Snippet: HK-2 cells were induced with LPS (10 μg/mL) (MCE, United States) combined with high glucose (30 mmol/L) to establish inflammation and apoptosis models ( ) and then treated with quercetin (Que) and finerenone (Fine) (MCE, United States, drugs were dissolved in DMSO, concentration 0.1%).

Techniques: CCK-8 Assay, Inhibition