Revealing potential anti-fibrotic mechanism of Ganxianfang formula based on RNA sequence

Abstract

Background Ganxianfang (GXF) formula as a traditional Chinese medicine (TCM) is used for liver fibrosis in clinical practice while its mechanism is unclear. The aim of this study is to explore the molecular mechanism of GXF against CCl4-induced liver fibrosis rats. Methods Detected the main compounds of GXF by UPLC-MS/MS. Evaluated the efficacy of GXF (1.58, 3.15, 4.73 g/kg/day) and Fuzheng Huayu (FZHY, positive control, 0.47 g/kg/day) through serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels and histopathological changes. Explored the underlying mechanisms by integrating our total liver RNA sequencing (RNA-seq) data with recent liver single-cell sequencing (scRNA-seq) studies. Verified potential pharmacodynamic substances of GXF by hepatic stellate cell (HSC)-T6 line. Results Main compounds were identified in GXF by UPLC-MS/MS, including baicalin, wogonoside and matrine etc. With GXF-high dose treatment, the elevation of ALT and AST induced by CCl4 were significantly reduced, and the protective effect of GXF-high dose treatment was better than FZHY. Liver histopathological changes were alleviated by GXF-high dose treatment, the ISHAK scoring showed the incidence of liver cirrhosis (F5/F6) decreased from 76.5 to 55.6%. The results of liver hydroxyproline content were consistent with the histopathological changes. RNA-seq analysis revealed the differential genes (DEGs) were mainly enriched in ECM-receptor interaction and chemokine signaling pathway. GXF effectively inhibited collagen deposition and significantly downregulated CCL2 to inhibit the recruitment of macrophages in liver tissue. Integrating scRNA-seq data revealed that GXF effectively inhibited the expansion of scar-associated Trem2+CD9+ macrophages subpopulation and PDGFRα+PDGFRβ+ scar-producing myofibroblasts in the damaged liver, and remodeled the fibrotic niche via regulation of ligand-receptor interactions including TGFβ/EGFR, PDGFB/PDGFRα, and TNFSF12/TNFRSF12a signaling. In vitro experiments demonstrated that baicalin, matrine and hesperidin in GXF inhibited the activation of hepatic stellate cells. Conclusions This study clarified the potential anti-fibrotic effects and molecular mechanism of GXF in CCl4-induced liver fibrosis rats, which deserves further promotion and application.

Expression profiles of circular RNAs and interaction networks of competing endogenous RNAs in neurogenic bladder of rats following suprasacral spinal cord injury

“…Evidence demonstrated that certain pathways play a role in developing organ fibrosis. For example, RNA-seq and single-cell sequencing were performed on rat liver tissues with CCl4-induced fibrosis, suggesting that fibrosis is closely related to the Chemokine signaling pathway ( Liu et al, 2022b ).…”

The Ethnopharmacological, Phytochemical, and Pharmacological Review of Euryale ferox Salisb.: A Chinese Medicine Food Homology

“…Euryale ferox seed (EFS) is a typical representative of "a medicine food homology species", as described in the Huangdi Nei Jing Tai Su (黄帝内经太素): "Eating it as food on an empty stomach, and taking it as medicine for patients" [1,2]. EFS is the dried seeds of the E. ferox Salisb.…”

Pharmacology, Toxicology, and Therapeutics of Minerals in Traditional Medicine 2021

“…Modulation of gut microbiota could be an important target for QSW [ 12 ] to produce neuroprotective effects but also for Hua-Feng-Dan to produce neuroprotective effects [ 17 ] fortifying the role of metal composition and herbal interactions in herbo-mineral preparations. RNA-Seq technology was also employed to analyze the adaptive mechanism of hepatoprotection with cinnabar and realgar-containing Hua-Feng-Dan [ 18 ] and antihepatic fibrotic mechanism with Ganxianfang formula [ 19 ].…”