Serum Metabolomics and Multiomics Validation Reveal Zishangan Granule-Induced P53-Bax/Casp3 Signaling in NSCLC.

Nonsmall cell lung cancer (NSCLC) remains a major cause of cancer mortality, underscoring the need for multitarget therapeutic candidates with defined mechanisms. This study investigated the anti-NSCLC activity of Zishangan Granule (ZSG), a four-herb formulation, using serum metabolomics, network pharmacology, molecular docking, public multiomics datasets, and experimental validation. Ultra-performance liquid chromatography-tandem mass spectrometry identified 15 serum-available constituents. Network analysis revealed 111 shared targets between these constituents and NSCLC-associated genes, with Tp53, Casp3, Bax, Pten, and Parp1 identified as core targets. Salvianolic acid L showed favorable predicted binding to these proteins, while public transcriptomic, proteomic, and immunohistochemical datasets supported their disease relevance. In a subcutaneous Lewis lung carcinoma (LLC) model, ZSG reduced tumor weight by 58.3% and increased tumor-cell apoptosis approximately threefold, accompanied by increased expression of P53, P21, Bax, Casp3, Pten, and Parp1. ZSG also reduced viability and colony formation and induced apoptotic morphology in LLC and A549 cells. These findings support the anti-NSCLC activity of ZSG through P53-associated Bax/Casp3 apoptotic signaling and warrant further preclinical evaluation.