Integrated transcriptomics and network pharmacology approaches to elucidate the anti-fibrotic mechanism of Eucommia ulmoides leaves against liver fibrosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Eucommia ulmoides Oliv. Leaves (EUL) are used as a traditional medicine. Eucommia ulmoides Oliv. is the only member of the Eucommiaceae family. Based on the nourish effect of EUL on liver meridian, the treatment mechanism of EUL on liver fibrosis (LF) was explored.

AIM OF THE STUDY: This study aims to investigate the effects and underlying mechanism of EUL on anti-fibrotic using in vitro and in vivo, and integrating transcriptomics and network pharmacology.

MATERIALS AND METHODS: TGF-β1 stimulated LX-2 cells were used to model fibrosis in vitro, while CCl4-induced mice served as an in vivo model. Cell activation, proliferation, migration, oxidative stress, and cytokine levels were assessed in vitro. The liver functions were evaluated by serum biochemical detection, histopathological analyses. Differentially expression genes (DEGs) between the control group and LF, LF group and EUL group were analyzed by transcriptomics analysis. Common targets were obtained by integrating transcriptomics and network pharmacology, and validated.

RESULTS: EUL suppressed HSCs activation, proliferation and migration in vitro. In CCl4 induced fibrotic mice, EUL reduced collagen deposition and improved liver function. Integrated analysis identified 66 common targets including AKT1, TNF, MMP9, CCND1, KDR and the PI3K-AKT as the central pathway.

CONCLUSIONS: The bioactive components of EUL ameliorate LF by inhibiting PI3K-AKT signaling pathway, thereby suppressing HSCs activation, proliferation, migration, and extracellular matrix deposition, while enhancing antioxidant capacity and attenuating inflammation. These findings elucidate the anti-fibrotic efficacy of EUL via PI3K-AKT inhibition, providing novel therapeutics for LF.