The mechanism of the water layer extract of Cichorium pumilum Jacq water extract in the treatment of Hepatic fibrosis was investigated using HPLC-MS/MS and network pharmacology.

Hepatic fibrosis (HF) represents a critical stage in the progression of chronic liver disease, for which effective treatments remain limited. Cichorium pumilum Jacq (CG), a traditional Chinese medicinal herb abundant in Xinjiang, is commonly used by the local Uyghur population for therapeutic purposes. Modern pharmacology indicates that CG possesses anti-fibrotic, anti-steatotic, anti-inflammatory, and antibacterial properties. However, the primary chemical constituents of Cichorium pumilum Jacq water extract (CGEF) and its anti-fibrotic mechanisms remain unclear. This study induced HF in SD rats using 40% carbon tetrachloride-olive oil and found that CGEF improved serum markers and mitigated hepatic pathological damage. CGEF components were identified via HPLC-MS/MS, with network pharmacology predicting key targets and enrichment analysis conducted. A total of 72 major compounds were identified, network pharmacology predicted 45 key targets, and enrichment results indicated CGEF may alleviate HF through a regulatory network centered on the TGF-β/Smad pathway. To validate this hypothesis, immunohistochemical analysis was performed on liver tissues from experimental animals. In vitro, TGF-β-induced HSC-T6 cell models were treated with CGEF to evaluate changes in TGF-β/Smad pathway-related gene expression. Immunohistochemical results indicated that CGEF significantly downregulated TGF-β1, TLR4, and α-SMA expression. In cell experiments, CGEF significantly reduced the gene expression of Smad2, Smad3, Smad7, TGF-β1, and PPP5C at 24 h. At 48 h, CGEF decreased the gene expression of Smad2, Smad3, TGF-β1, and PPP5C while increasing the gene expression of Smad7. Thus, CGEF may alleviate HF primarily by regulating the TGF-β/Smad pathway.