Targeting the p38/MAPK pathway to induce apoptosis: a multidimensional mechanistic exploration of Mentha and its active compound diosmetin against liver cancer

Liver cancer, a malignancy with high global incidence and mortality, currently relies on surgical resection, radiotherapy, and chemotherapy, all of which face significant limitations, necessitating novel therapeutic strategies. Mentha (ME), a medicinal and edible herb, has demonstrated antioxidant, anti-inflammatory, and broad-spectrum anticancer activities, yet its molecular mechanisms against liver cancer remain unclear. This study will comprehensively explore the anti-liver cancer mechanisms of ME and its key bioactive constituent, diosmetin (Dio). A multi-disciplinary approach, which incorporates network pharmacology, molecular docking, and molecular dynamics simulations, was adopted in this study to thoroughly explore the bioactive components of ME and the mechanisms through which they exert anti-liver cancer effects. Functional validation was conducted through CCK-8 viability assays, clonogenic survival assays, scratch wound healing, Transwell migration assays, Western blotting, immunofluorescence, and TUNEL apoptosis assays in human liver cancer cell lines (HepG2 and HuH-7). ME exhibited potent anti-Liver cancer activity, significantly suppressing cell viability, proliferation (CCK-8/clonogenic assays), and migration (scratch/Transwell assays, P < 0.01), while downregulating metastasis-related proteins MMP2/MMP9 (Western blot/immunofluorescence, P < 0.01). Network pharmacology identified TP53 (p53), TNF, CASP3 (caspase3), IL6, and IL1B as core targets. Based on the results of molecular docking (ΔG < - 4 kcal/mol) and molecular dynamics simulations (maximum ΔTotal), Dio was prioritized for subsequent experimental validation. Further validation demonstrated Dio's multi-modal efficacy: GO/KEGG analysis revealed its dual action via p38/MAPK signaling and apoptosis pathways, corroborated by upregulated pro-apoptotic markers (p53, caspase3, Bax, p38) and downregulated Bcl2 (P < 0.01), alongside TUNEL-confirmed apoptosis induction (P < 0.01). This study is the first to demonstrate that ME and its active compound Dio inhibit liver cancer progression via multi-target regulation of the p38/MAPK pathway, providing a theoretical foundation for developing ME-based natural therapeutics against liver cancer.