Notoginsenoside R2 attenuates hepatic fibrosis via STAT3-dependent hepatic stellate cells senescence induction and inflammatory microenvironment suppression

Hepatic fibrosis (HF) continues to be a significant global health concern, substantially contributing to morbidity and mortality due to the absence of effective therapeutic options. This study examines the pharmacological effectiveness and underlying mechanisms of Notoginsenoside R2 (R2) in mitigating HF, aiming to find a new multifunctional candidate for therapeutic application. An integrative methodology utilizing network pharmacology, molecular docking, and experimental validation was implemented. In vitro models (HSC-T6), in vivo systems (zebrafish), and microinjection of morpholinos were employed to corroborate the antifibrotic effects of R2 and transcription 3 (STAT3)-dependent processes. Network pharmacology identified 32 common targets between R2 and HF, with a particular emphasis on pathways critical for the activation of HSCs. Molecular docking confirmed strong interactions between R2 and signal transducer and activator of STAT3. In vitro , R2 inhibited HSCs proliferation and decreased the expression of α-SMA, COL-I, Desimin and TIMP1. In vivo , R2 mitigated thioacetamide-induced fibrosis in zebrafish, leading to decreased collagen deposition and suppression of pro-inflammatory cytokines. Mechanistically, R2 induced senescence in HSCs via the STAT3 pathway, characterized by increased expression of cyclin-dependent kinase inhibitor 2A (CDKN2A/p16) and cyclin-dependent kinase inhibitor 1A (CDKN1A/p21), as well as components of the senescence-associated secretory phenotypes (SASPs). This study identified R2 as a regulator of STAT3 with dual antifibrotic effects: reduction of the inflammatory microenvironment and induction of senescence. These findings position R2 as a viable treatment candidate for HF, necessitating additional clinical investigation.