Berberine protects against hepatocellular carcinoma progression by regulating intrahepatic T cell heterogeneity

Accumulating evidence suggests that berberine (BBR) exhibits anti-cancer effects in hepatocellular carcinoma (HCC). However, the mechanisms by which BBR regulates the immunological microenvironment in HCC has not been fully elucidated. In this study, we established a mouse model of orthotopic HCC and treated with varying doses of BBR. BBR showed effectiveness in reducing tumor burden in mice with HCC. Cytometry by time-of-flight depicted the alterations in the tumor immune landscape following BBR treatment, revealing the enhancement in the T lymphocytes effector function. In particular, BBR decreased the proportion of TCRbhiPD-1hiCD69+CD27+CD8a+ effector CD8+ T lymphocytes and increased the proportion of Ly6ChiTCRb+CD69+CD27+CD62L+ central memory CD8+ T lymphocytes. Single-cell RNA sequencing further elucidate the effects of BBR on transcriptional profiles of liver immune cell and confirmed the phenotypical heterogeneity of T lymphocytes in HCC immune microenvironment. Additionally, we found that BBR potentially regulated the antitumor immunity in HCC by modulating the receptor-ligand interaction among immune cells mediated by cytokines. In summary, our findings improve the understanding of BBR's impact on protecting against HCC, emphasizing BBR's role in regulating intrahepatic T cell heterogeneity. BBR has the potential to be a promising therapeutic strategy to hinder the advancement of HCC. The HCC mouse models were randomly divided into three groups: HCC mouse models treated with PBC (HC group), HCC mouse models treated with low-dose BBR (LB group), HCC mouse model treated with high-dose BBR (HB group). BBR with a purity of 98% was provided by Meilun Biology (Dalian, China) and administered intraperitoneally to the HCC mouse model. The HB group received 200 μL of a BBR solution at a dosage of 30 mg/kg, while the LB group received 10 mg/kg. The HC group was given an equivalent volume of phosphate buffer saline (PBS). Administration was carried out every other day for a period of 14 days. Cytometry by time-of-flight depicted the alterations in the tumor immune landscape following BBR treatment.Single-cell RNA sequencing further elucidate the effects of BBR on transcriptional profiles of liver immune cell and confirmed the phenotypical heterogeneity of T lymphocytes in HCC immune microenvironment.