六味五灵片（LWWL）是经中国国家药品监督管理局（NMPA）批准的中成药。临床上，它用于治疗肝细胞癌（HCC）之前的一系列肝脏疾病，包括肝炎、肝纤维化和肝硬化。 LWWL被认为可以抑制HCC的炎症转化，这可能对HCC的预防和治疗产生积极影响。然而，其确切的作用机制仍不清楚。为了研究LWWL如何有效治疗HCC并验证该过程中涉及的通路，建立二乙基亚硝胺（DEN）诱导的HCC体内模型来研究LWWL的作用关于 HCC 的发展。分析大鼠血清的天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、碱性磷酸酶（ALP）和γ-谷氨酰转肽酶（GGT）。大鼠肝组织用苏木精和伊红（H

LiuweiWuling Tablet (LWWL) is a patented Chinese medicine approved by the Chinese National Medical Products Administration (NMPA). Clinically, it is used to treat a range of liver diseases that precede hepatocellular carcinoma (HCC), including hepatitis, liver fibrosis and cirrhosis. LWWL is hypothesized to inhibit the inflammatory transformation of HCC, which may have a positive impact on the prevention and treatment of HCC. However, its exact mechanism of action remains unknown.To investigate how LWWL is effective in the treatment of HCC and to validate the pathways involved in this process.In vivo model of HCC induced by diethylnitrosamine (DEN) was established to study the effect of LWWL on the development of HCC. The rat serum was analyzed for aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transpeptidase (GGT). The rat liver tissues were stained with hematoxylin and eosin (H&E) and Masson's trichrome for pathological analysis. Rat liver tissue was subjected to transcriptome sequencing. Expression of inflammatory and liver fibrosis-related factors in bone marrow-derived macrophages (BMDMs) and LX-2 cells was detected by QPCR, ELISA and Western blot (WB). The expression of apoptosis and stemness genes in HepG2 and Huh7 cells was assessed by LWWL through flow cytometry and QPCR. Transcriptomics, network pharmacology, WB, and QPCR were employed to validate the mechanisms associated with the amelioration of HCC development by LWWL.LWWL significantly reduced the severity of hepatitis and liver fibrosis, the expression of tumor stemness genes, and a significant decrease in the incidence of HCC. In addition, LWWL inhibited the release of inflammatory substances and nuclear accumulation of P65 protein in BMDMs as well as the conversion of LX-2 cells to fibroblasts. LWWL inhibited the proliferation of HepG2 and Huh7 cells, including the initiation of apoptosis and the reduction of stemness gene expression. Importantly, LWWL regulates the PI3K/AKT/NF-κB pathway, which affects hepatic inflammation and cancer progression.LWWL inhibited the occurrence and development of HCC by modulating the severity of hepatitis and liver fibrosis, indicating the potential clinical relevance of LWWL in preventing and treating HCC.Copyright © 2023. Published by Elsevier B.V.