Rutaecarpine alleviates liver fibrosis by modulating the TRPV1/Connexin 43 axis: A study based on transcriptome data, single-cell sequencing data, and cell experiments.

Liver fibrosis is a major pathological process leading to cirrhosis and liver failure, posing a significant global health challenge. Rutaecarpine has potential benefits in liver disease and is an agonist of TRPV1. This study aimed to explore the potential mechanism by which rutaecarpine alleviates liver fibrosis. Based on the GSE84044 dataset, correlation analysis and logistic regression were performed to evaluate the relationships among TRPV1, connexin 43 (Cx43), and liver fibrosis. The diagnostic performance of TRPV1, Cx43, and their combined gene model for liver fibrosis was assessed using receiver operating characteristic (ROC) curve. Interaction and mediation analyses were conducted to explore the role of Cx43 in TRPV1-liver fibrosis. Single-cell sequencing analysis investigated cellular interactions and their associations with TRPV1 and Cx43. Molecular docking was used to assess the binding affinity of rutaecarpine with TRPV1 and Cx43. A lipopolysaccharide (LPS)-induced fibrosis cell model was established for validation. TRPV1 expression was negatively correlated with Cx43. Low TRPV1 and high Cx43 levels were significantly associated with increased risk of liver fibrosis (OR = 8.500, 95 % CI: 2.457-35.349). The TRPV1-Cx43 gene model effectively predicted liver fibrosis. Additionally, TRPV1 was mainly expressed in hepatocytes, while Cx43 was expressed in endothelial cells. Rutaecarpine stably interacted with TRPV1 and Cx43. Validation experiments confirmed that rutaecarpine reversed LPS-induced fibrotic changes and modulated TRPV1 and Cx43 expression in a dose-dependent manner. TRPV1 negatively regulated Cx43 in liver fibrosis, and rutaecarpine alleviated liver fibrosis by modulating the TRPV1/Cx43 axis.