Single-cell transcriptomics reveals the ameliorative effect of Gastrodin on cholestatic liver fibrosis

Cholestatic liver fibrosis represents a significant clinical challenge with limited therapeutic options. This study examines the therapeutic potential and underlying mechanisms of Gastrodin (GAS), a primary bioactive compound derived from Gastrodia elata, in the treatment of cholestatic liver fibrosis. Utilizing two established murine models (DDC-diet and BDL), we demonstrated that GAS administration significantly alleviated liver injury, reduced serum bilirubin and bile acids, attenuated collagen deposition, and improved survival. Through an integrated analysis of single-nucleus RNA sequencing (snRNA-seq) data from our study and a publicly available dataset, we delineated the cellular and molecular landscape of GAS action. We found that GAS facilitated hepatocyte repair by reversing the expression of key sulfotransferases Sult2a1 and Sult1e1, enhancing detoxification. Concurrently, GAS markedly reduced the proportion of pro-inflammatory Kupffer cells and downregulated their expression of inflammatory mediators (e.g., TNF-α, Il-6), potentially through upregulating immunomodulatory genes (Pilrb1, Ifi27l2a and Rtp4). Furthermore, GAS modulated hepatic stellate cell (HSC) heterogeneity, expanding a novel Serpina⁺ HSC subpopulation with an anti-fibrotic signature. Our findings reveal that GAS ameliorates cholestatic liver fibrosis through multi-faceted mechanisms targeting hepatocytes, Kupffer cells, and HSCs, positioning it as a promising natural therapeutic agent and highlighting Sult2a1, Sult1e1, and Serpina1a as potential therapeutic targets.