Huangkui capsule combined with finerenone attenuate diabetic n ephropathy by regulating JAK2/STAT3 signaling pathway based on network pharmacology, molecular docking and experimental verification

Xunake Liua^*Chunjiang ZhangaYanjie FuaJingJing DaiaJiaying LuaGang LuaXiaoping Yanga

• First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi, China

Introduction: Diabetic nephropathy (DN) is a serious complication of diabetes with limited therapeutic opt ions. While Huangkui capsule (HKC) and finerenone individually show potential in DN management, their combined mechanism remains unclear. This study aimed to explore the therapeutic effects and underlying mechanisms of HKC combined with finerenone for DN.Methods: An integrative approach combining network pharmacology, molecular docking, kinetic simulation s, and experimental validation was employed. Active components of HKC and finerenone, along with thei r potential targets, were identified through database mining. A "drug-component-target-disease" network wa s constructed, and interactions with the JAK2/STAT3 pathway were validated. In vivo, DN mice were tre ated with HKC, finerenone, and their combination (CDI group), while in vitro, HK2 cells were treated w ith quercetin (a core HKC component) and finerenone.The binding index between quercetin and finerenon e was analyzed by Chou-Talalay method.Results: Network pharmacology identified three core HKC components (quercetin, myricetin, gossypetin) a nd 11 key targets (e.g., JAK2, STAT3, AKT1). Molecular docking revealed strong binding affinity betwee n quercetin-finerenone and JAK2/STAT3 (ΔG = -65.465 kcal/mol for STAT3-quercetin). In DN mice, com bined therapy significantly reduced 24-hour urinary protein (358.54±21.21 mg/L vs. 1046.48±72.84 mg/L i n the model group, p < 0.001), improved serum creatinine/urea nitrogen, and downregulated IL-6/TNF-α. It also suppressed pro-apoptotic genes (Bax, Caspase3/8, PARP) while upregulating Bcl-2. Histopathology s howed reduced tubular injury markers (NGAL, KIM-1) and fibrosis (p<0.05). In HK2 cells, quercetin+fine renone synergistically inhibited apoptosis and Inflammation(p<0.05), and the combined index (cl) was calc ulated to be Less than 1. STAT3 overexpression exacerbated inflammation/apoptosis, which was reversed by combined treatment (p<0.01).Conclusions: HKC combined with finerenone mitigates DN progression by inhibiting the JAK2/STAT3 pat hway, reducing inflammation, apoptosis, and tubular injury. These findings provide a mechanistic basis for clinical application of this combination therapy.