Single-cell Transcriptome Analysis Reveals the Potential Role of Hepatic Stellate Cells in Liver Fibrosis.

Abstract

Background: Upon activation, hepatic stellate cells (HSCs) can convert into fibroblasts and increase the production of extracellular matrix, a major cause of liver fibrosis (LF) and a growing health issue worldwide. Other mechanisms by which HSCs may induce fibrosis remain to be explored, and the role of cell dynamic gene expression in liver fibrogenesis is not well understood. In this study, analysis by single-cell transcriptome sequencing (scRNA-seq) was used to explore the potential effects of HSCs in a bile duct ligation (BDL)-induced mouse model of LF, followed by the identification of novel targets for clinical diagnosis.

Methods: Liver tissue collected from BDL and sham-operated C57BL/6J mice was used for scRNA-seq. To systematically dissect the molecular and cellular events following fibrosis, the scRNA-seq data was analyzed for differential gene expression, KEGG, pseudotime trajectory, and cellular communication. Morphological changes in the BDL and sham livers were examined by hematoxylin and eosin (H&E) staining, Masson's trichrome staining, fiber staining, and Sirius red staining.

Results: The scRNA-seq analysis performed on the BDL and sham groups revealed the gene expression of 20,764 cells across 27 cell types. Antioxidant levels declined markedly in HSCs from BDL mice, leading to a more pronounced occurrence of ferroptosis. We also found evidence suggesting that elevated apelin signaling and platelet activation in HSCs contributed to the increased synthesis of extracellular matrix and collagen fibers. The large accumulation of immune cells in the liver of BDL mice induces different outcomes for HSCs.

Conclusion: The results of this study provide further insight into the cellular and molecular alterations that occur within a specific subset of HSCs during LF, offering valuable information on potential targets for therapeutic intervention.