Mechanistic insights into Tuoli Xiaozheng Formula for hepatocellular carcinoma: An integrated network pharmacology, molecular docking, and Mendelian randomization analysis

Hepatocellular carcinoma (HCC) is a malignancy with a poor prognosis and low overall survival. Tuoli Xiaozheng Formula (TLXZF), a traditional Chinese compound prescription, has shown clinical efficacy in treating HCC, but its multi-component and multi-target mechanisms remain unclear. This study aimed to elucidate the molecular mechanisms of TLXZF against HCC by integrating network pharmacology, molecular docking, and Mendelian randomization (MR) analyses. Active compounds and targets were identified from the Traditional Chinese Medicine Systems Pharmacology, high-throughput experiment- and reference-guided database of traditional Chinese medicine, and SwissTargetPrediction databases, while HCC-related targets were obtained from Online Mendelian Inheritance in Man, GeneCards, DrugBank, and therapeutic target database. The intersection yielded 227 overlapping targets. Network analysis revealed quercetin, kaempferol, stigmasterol, luteolin, and isorhamnetin as core bioactive components, and protein–protein interaction network analysis identified TP53, AKT1, CTNNB1, EGFR, IL-6, STAT3, TNF, BCL-2, and CASP3 as key targets. Kyoto Encyclopedia of Genes and Genomes enrichment indicated that TLXZF mainly acts through PI3K-AKT, MAPK, and HIF-1 signaling pathways. Molecular docking confirmed stable binding between core compounds and targets. MR analysis indicated that genetically predicted AKT1 levels were modestly associated with a reduced risk of HCC (odds ratio = 0.9986, P  = .000758), although the effect size was close to the null, suggesting a weak but statistically detectable lifelong genetic association. Overall, this integrative analysis suggests that TLXZF may exert its effects on HCC through a synergistic “multi-component-multi-target-multi-pathway” regulatory pattern, primarily involving oncogenic, inflammatory, and survival-related signaling networks. These findings are hypothesis-generating and provide an initial basis for the identification and screening of core bioactive components, key targets, and relevant pathways to support future experimental validation and clinical research.