The Mechanism of Huaiqihuang in the Treatment of Nephrotic Syndrome: An Integrated Study Based on Network Pharmacology, Molecular Docking, Molecular Dynamics Simulation, and Experimental Validation.

INTRODUCTION

Huaiqihuang (HQH) is a traditional Chinese medicine commonly used in the clinical management of nephrotic syndrome (NS). Nevertheless, the molecular mechanisms underlying its therapeutic efficacy remain unclear. This study aimed to elucidate the molecular basis of HQH in NS treatment through an integrated approach involving network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation.

METHODS

Active compounds and their potential targets associated with HQH were identified using the TCMSP and HERB databases. NS-related targets were retrieved from the GeneCards, OMIM, and DisGeNET databases. Protein-protein interaction (PPI) networks were constructed using Cytoscape to identify key targets, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Molecular docking and molecular dynamics simulations were performed to evaluate the binding affinity and stability of interactions between core compounds and targets. We accessed all databases and collected the data in July 2024. In vitro experiments were conducted using a doxorubicin (DOX)-induced MPC5 podocyte injury model. The effects of HQH on inflammatory mediators, oxidative stress, and related signaling pathways were assessed using CCK-8 assays, immunofluorescence, RT-qPCR, and Western blotting.

RESULTS

Network pharmacology analysis identified zhonghualiaoine 1, physalin A, and methylluteolin as major bioactive components of HQH, with ALB, AKT1, and EGFR emerging as core targets. GO and KEGG enrichment analyses indicated that HQH may exert therapeutic effects by regulating oxidative stress and inflammatory responses, primarily through the PI3K/AKT signaling pathway. In vitro experiments demonstrated that HQH significantly improved podocyte viability following DOX-induced injury and markedly reduced NLRP3 inflammasome activation. Mechanistically, HQH promoted the phosphorylation of PI3K and AKT and facilitated the expression of Nrf2, thereby enhancing antioxidant capacity in podocytes.

DISCUSSION: This study confirms that Huangqi Huang (HQH) effectively alleviates inflammatory responses and oxidative stress damage in podocytes in vitro, with its mechanism potentially involving the regulation of multiple pathways such as PI3K-AKT, as predicted by network pharmacology. In the experiment, HQH demonstrated particularly potent inhibitory effects on NLRP3 inflammasomes, suggesting anti-inflammatory action may be a key mechanism underlying its renal protective effects. Concurrently, activation of the PI3K/AKT/Nrf2 antioxidant pathway likely constitutes a synergistic mechanism for interrupting the vicious cycle of injury. Although the roles of NLRP3 and the PI3K/AKT pathway in nephrotic syndrome have been previously reported, this study is the first to reveal, in a podocyte model, that these two pathways may serve as core target axes for HQH intervention. This finding provides a new perspective for elucidating the modern biological implications of its therapeutic mechanisms.

CONCLUSION

This study provides systematic evidence that HQH exerts renoprotective effects in nephrotic syndrome through multi-component and multi-target mechanisms, predominantly involving activation of the PI3K/AKT/Nrf2 signaling pathway and inhibition of NLRP3 inflammasome-mediated inflammation.