Background: Chronic nonbacterial prostatitis (CNP), the major subset of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), imposes a substantial global burden yet lacks satisfactory therapies. Maizibizi Wan (MZBZ) has long been used clinically for prostatitis, but its pharmacodynamic substance basis and mechanisms remain unclear. Methods: Ultra-high-performance liquid chromatography–Q-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) coupled with Global Natural Products Social Molecular Networking (GNPS) molecular networking profiled MZBZ constituents and rat plasma–exposed prototype components and metabolites was used. Based on blood-absorbable components, network pharmacology predicted core targets/pathways; representative interactions were validated by molecular docking. A λ-carrageenan–induced CNBP rat model underwent histopathology (H&E), serum cytokine assays (TNF-α, IL-1β, IL-6/IL-17), immunohistochemistry (COX-2, TNF-α, MMP-9), and Western blotting (P-p65/p65, p-AKT/AKT, COX-2, TGF-β1, BCL2). Results: A total of 188 chemical constituents were identified in MZBZ (79 flavonoids, 38 organic acids, 30 alkaloids, 15 phenylpropanoids, 7 steroids, 4 phenylethanoid glycosides, 15 others). A total of 35 blood-absorbable components (18 prototype components, 17 metabolites) were identified, mainly involving Phase I oxidation and Phase II glucuronidation/sulfation. Network analysis yielded 54 core targets enriched in NF-κB and PI3K/AKT signaling and apoptosis. Docking indicated stable binding of key flavonoids to COX-2, NFKB1, TNF, IL-6, and BCL2. In vivo, MZBZ ameliorated prostatic inflammation, reduced serum TNF-α/IL-1β/IL-6/IL-17 (p < 0.05 or p < 0.01); decreased P-p65/p65, p-AKT/AKT, COX-2, and TGF-β1; and increased BCL2 in prostate tissue. Conclusions: MZBZ exerts anti-CNBP effects via multi-component synergy (prototypes + metabolites) that suppresses inflammatory cytokines, modulates apoptosis, and inhibits NF-κB and PI3K/AKT pathways. These findings provide a mechanistic basis and quality control cues for the rational clinical use of MZBZ.

Cigarette smoke (CS) is a leading cause of respiratory diseases and cellular damage. Apigenin (Api), a natural flavonoid, exhibits antioxidant and anti-inflammatory properties, suggesting a potential protective role against cigarette smoke extract (CSE)-induced cellular damage. Aim: This study aimed to investigate the protective effects of Api on Beas-2B cells exposed to CSE. Methods: We optimized experimental conditions by treating Beas-2B cells with varying concentrations of Api (0 to 200[ ] [ ]mol/L) and CSE (1–30%) to determine the optimal concentrations and exposure times using the MTT assay for cell viability. Cells were then pretreated with 25, 50, and 100[ ] [ ]mol/L Api for 24[ ]h before a 4-h exposure to 5% CSE. The protective effects of Api were evaluated through multiple assays: cell viability (MTT), apoptosis and cell cycle analysis (flow cytometry), mitochondrial morphology (scanning electron microscopy), mitochondrial membrane potential and ATP content (TMRE), superoxide dismutase (SOD) activity (WST-8), and malondialdehyde (MDA) level (TBA). To elucidate the molecular mechanisms, we performed RNA sequencing to identify differentially expressed genes (DEGs) and conducted bioinformatics analysis. The PI3K/AKT and mitogen-activated protein kinase (MAPK) signaling pathways were confirmed using Western blot. Results: Api significantly increased Beas-2B cell survival within the tested concentration range ([ ]). CSE treatment for 4[ ]h significantly inhibited cell growth ([ ]), which was mitigated by Api pretreatment ([ ]), reducing apoptosis and modulating the cell cycle. Api also reduced MDA levels and enhanced SOD activity in CSE-treated cells ([ ]). RNA sequencing revealed 1761 DEGs in the Api-treated group compared to the CSE-only group, with 742 upregulated and 1019 downregulated genes, primarily associated with the MAPK and PI3K/AKT pathways. qPCR validation confirmed the top 10 upregulated and downregulated genes. Importantly, 50[ ] [ ]mol/L Api downregulated the protein expression of p-Akt, p-ERK, and p-MAPK in CSE-treated cells ([ ]). Conclusion: Our findings demonstrate that Api significantly reduces CSE-induced damage in Beas-2B cells, potentially through the inhibition of PI3K/AKT and MAPK signaling pathways and the regulation of multiple genes and mitochondrial function.