Biejiajian pill (BJJP) is a canonical formula that is clinically used to treat chronic liver disease, especially to decrease the incidence of hepatocellular carcinoma (HCC). However, the mechanisms underlying the prevention of HCC progression by BJJP remain unclear. This study aimed to determine whether BJJP inhibits HCC progression by downregulating platelet-derived growth factor receptor beta (PDGFRβ) signaling in cancer-associated fibroblasts (CAFs) in a mouse model of diethylnitrosamine (DEN)/carbon tetrachloride (CCl₄)-induced HCC. C57BL/6 male mice were intraperitoneally injected with DEN 2 weeks after birth, followed by repeated injections of CCl₄ weekly from 6 weeks of age onwards, to recapitulate features of HCC. At week 14, BJJP was orally administered to mice. The effects of BJJP on HCC progression were evaluated using histology, immunohistochemistry, and serum biochemical marker levels. Transcriptome analysis, molecular docking, quantitative real-time PCR, and Western blot were used to study the genes targeted by BJJP and the associated signaling pathway. The effects of BJJP on PDGFRβ signaling in CAFs and the underlying mechanism were demonstrated. BJJP treatment significantly suppressed carcinogenesis and cancer progression, and it ameliorated liver inflammation in mice with HCC. A total of 176 genes, including PDGFRβ, were significantly downregulated after BJJP treatment and five components of BJJP with high binding affinity to PDGFRβ were identified. BJJP inhibited the phosphorylation of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and glycogen synthase kinase 3 beta (GSK3β) by suppressing PDGFRβ expression in CAFs, and it also downregulated the expression of the downstream proteins hepatocyte growth factor (HGF) and vascular endothelial growth factor A (VEGF-A). Furthermore, BJJP-containing serum consistently reduced PDGFRβ, HGF, and VEGF-A expression levels in HSC-derived CAFs in vitro. Importantly, PDGF-BB induced PDGFRβ activation in CAFs and both BJJP and sunitinib (a kinase inhibitor) inhibited PDGF-BB/PDGFRβ signaling. BJJP inhibits the progression of HCC through suppressing VEGF-A and HGF expression in CAFs by downregulating PDGFRβ signaling.

The efficacy of immunotherapy for hepatocellular carcinoma (HCC) is often limited by an immunosuppressive tumor microenvironment (TME), largely driven by tumor metabolic reprogramming (the Warburg effect) that impairs CD8⁺ T cells. Targeting this metabolism to restore immunity is a promising strategy. Bie-Jia-Jian Pill (BJJP), a traditional formula for "softening hardness and dissipating masses," conceptually aligns with modulating the TME. However, its role in reprogramming tumor metabolism to enhance immunotherapy remains unclear. We aimed to investigate if BJJP enhances anti-tumor immunity by inhibiting the Warburg effect, elucidate its molecular mechanism, and evaluate its synergy with PD-L1 blockade. Using UHPLC-MS, the major components of BJJP were analyzed and its efficacy was evaluated in combination with the PD-L1 neutralizing antibody for HCC treatment. CD8⁺ T cells and tumor markers were assessed in vitro and in vivo using flow cytometry. Transcriptomics, metabolomics, Seahorse metabolic assays, Western blotting, ELISA, and immunofluorescence staining were employed to identify and validate the mechanisms and targets of BJJP. Its targeting capabilities were confirmed by lentivirus and siRNA transfections, network pharmacology and molecular docking. BJJP suppressed HCC cell proliferation, growth and migration, promoted their apoptosis, reduced PD-L1 expression, and enhanced CD8⁺ T cell cytotoxicity and infiltration in the tumor. BJJP and immunotherapy exerted synergistic effects, including inhibiting tumor cell proliferation, increasing immune cell infiltration, and improving treatment efficacy. BJJP suppressed glycolysis in HCC cells, alleviated the excessive uptake of energy substrates, and reduced lactate accumulation in the microenvironment, thereby mitigating CD8⁺ T cell exhaustion. This glycolytic metabolic reprogramming was related to BJJP's effect on inhibiting the HIF-1α-PI3K/AKT/mTOR axis. Additionally, BJJP inhibited the downstream NF-κB activation, down-regulated CCL20, and alleviated the inhibitory effect of CCL20 against CD8⁺ T cells. BJJP targets the HIF-1α-PI3K/AKT/mTOR axis to reprogram tumor glycolysis, reduce CCL20 secretion, enhance CD8⁺ T cell immune response, and modulate the tumor microenvironment, thereby exerting anti-tumor effects against HCC. When used in combination with immunotherapy, BJJP can improve treatment efficacy, providing valuable evidence for clinical treatment.

Nephrotoxicity is the most common complication of cisplatin, and effective therapeutic strategies are still limited. Vascular dysfunction is thought to be early response after cisplatin administration and contributes to renal damage. However, the precise mechanisms underlying this process have yet to be fully elucidated. In this study, we observed a significant increase in renal vascular permeability following cisplatin administration in vivo, accompanied by a rapid loss of perivascular pericytes. Consistently, in vitro experiments revealed that cisplatin markedly accelerated cell death in primary human kidney microvascular pericytes (HKMPs). RNA-sequencing and proteomics analysis further identified a significant downregulation of six-transmembrane epithelial antigen of the prostate 4 (STEAP4) mRNA and protein expression in HKMPs after cisplatin exposure. Mechanistically, reduced STEAP4 expression facilitates iron accumulation and lipid peroxidation in HKMPs, leading to potentiated ferroptosis, whereas STEAP4 overexpression conferred protection against cisplatin-induced pericytes injury. Finally, targeted delivery of AAV9^-STEAP4 via renal artery injection efficiently increased renal STEAP4 expression, reduced pericyte loss, and attenuated renal vascular leakage. Furthermore, the AAV9^-STEAP4 group exhibited significantly decreased kidney injury compared to the AAV9^-NC group. Collectively, these findings demonstrate that STEAP4 mediates pericyte injury in renal vascular dysfunction and suggest that targeting STEAP4 could represent a novel therapeutic strategy for cisplatin-induced acute kidney injury (AKI).