Ginsenosides alleviate the progression of hepatic fibrosis by targeting the liver circadian clock gene CLOCK

Hepatic fibrosis (HF) serves as a critical pathological process underlying the progression of cirrhosis and hepatocellular carcinoma. Despite its clinical relevance, effective anti‑fibrotic therapies remain scarce. Although total ginsenosides (GSS) can ameliorate hepatic oxidative damage and attenuate the development of HF, the molecular mechanism mediating their anti‑fibrotic effects remains incompletely elucidated. Notably, circadian clock gene dysregulation is associated with hepatic metabolic dyshomeostasis and the fibrotic process. As a core circadian gene in the liver, clock circadian regulator (CLOCK) serves a role in the progression of fibrosis. The present study hypothesized that GSS may exert anti‑fibrotic effects by restoring hepatic circadian rhythmicity through circadian clock‑dependent pathways. Using a well‑established carbon tetrachloride‑induced murine model of HF in C57BL/6 mice, GSS was administered intraperitoneally at graded doses of 50, 100 and 200 mg/kg. Subsequently, serum and liver tissues were collected from mice for histopathological examination. Furthermore, the expression levels of fibrotic markers (α‑smooth muscle actin and collagen type 1) and circadian rhythm genes [CLOCK, basic helix‑loop‑helix ARNT‑like 1, cryptochrome (CRY)1, CRY2, period circadian protein homolog (PER)1 and PER2 were assessed. At the cellular level, LX‑2 hepatic stellate cells (HSCs) were activated with TGF‑β1 and treated with GSS, after which, RNA sequencing was performed on GSS‑treated cells. In addition, the CLOCK inhibitor (CLK8) was used for in vitro experiments. The results of the present study revealed that GSS intervention effectively reversed the dysregulation of circadian gene expression and alleviated the progression of fibrosis. Molecular docking analysis further supported the interaction between GSS active components and the CLOCK protein. Thus, the current study focused on GSS and explored its role in ameliorating HF through regulation of the core circadian gene CLOCK. Collectively, the findings demonstrated that GSS may alleviate HF by rescuing circadian clock‑driven circadian homeostasis in hepatocytes and HSCs, thereby providing mechanistic insight into traditional Chinese medicine‑based interventions and identifying CLOCK as a druggable node for fibrosis therapy.