Work

Skullcapflavone II induces G2/M phase arrest in hepatic stellate cells and suppresses hepatic fibrosis

Year: 2025

Type: article

Abstract: This investigation explored the therapeutic effects and mechanisms of Skullcapflavone II in hepatic fibrosis (HF). The optimal concentration of Skullcapflavone II for LX2 hepatic stellate cells was determined using the CCK8 assay. EdU staining and flow cytometry were utilised to assess cell proliferation and G2/M phase arrest. Mice with carbon tetrachloride-triggered HF were administered Skullcapflavone II at low (15 mg/day), medium (30 mg/day), and high (60 mg/day) doses. Subsequently, hepatic damage and fibrosis were assessed via body weight, liver index, biochemical markers, and histopathological staining. Immunohistochemistry for Collagen I and α-SMA were utilised to examine hepatic stellate cell (HSC) activation. RNA sequencing was utilised to ascertain differentially expressed genes. Molecular docking simulated interactions among Skullcapflavone II and target proteins as well as outcomes were validated by implementing western blotting, immunohistochemistry, and RT-qPCR. Skullcapflavone II inhibited LX2 cell proliferation and triggered G2/M phase arrest. Its optimal intervention concentration was 160 μM. In vivo , it ameliorated hepatic function, diminished serum indicators of fibrosis, and suppressed HSC activation. Diminished collagen sediment was validated utilising histopathological examination, whereas immunohistochemistry indicated decreased expression of Collagen I and α-SMA. Additionally, molecular docking showed strong binding of Skullcapflavone II to DNA replication-related proteins. Western blotting and RT-qPCR implied that Skullcapflavone II disrupted DNA replication, which triggered G2/M arrest and hindered HSCs activation and proliferation. The abovementioned mechanisms of action of Skullcapflavone II substantiate its prospective clinical application against HF. • Skullcapflavone II, from Scutellaria baicalensis, induces G2/M arrest in hepatic stellate cells, thus inhibiting their activation and proliferation, which are key factors in hepatic fibrosis progression. • Skullcapflavone II shows anti-fibrotic effects in vivo by improving liver function and reducing serum liver fibrosis markers in a carbon tetrachloride-induced mouse model. • The therapeutic mechanism involves disrupting DNA replication in hepatic stellate cells, leading to G2/M arrest and decreased cell activation. • Through RNA sequencing and molecular docking, its interaction with key proteins in DNA replication has been revealed, offering insights for new therapies against hepatic fibrosis.

Source: European Journal of Pharmacology

DOI: 10.1016/j.ejphar.2025.177522

Authors: Guoyi Rong, Jun Liu, Yunheng Yang, Shang Wang, Wenfu Cao

Institutions: Chongqing Medical University, Army Medical University, The Affiliated Yongchuan Hospital of Chongqing Medical University

Language: English

Topic: Liver physiology and pathology

Subfield: Hepatology

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,230