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.