Comprehensive two-dimensional primary hepatic stellate cell (HSC) membrane chromatography-based screening of anti-fibrotic components from Schisandra chinensis (Turcz.) Baill. and mechanistic insights into gomisin N against liver fibrosis

Abstract

Ethnopharmacological relevance: Schisandra chinensis, a renowned medicinal plant in traditional Chinese, Korean, and Russian ethnomedicine, has been widely used for its hepatoprotective, adaptogenic, and tonic properties. In traditional Chinese medicine, it is classified as a superior herb known to astringe essence, nourish the kidneys, and calm the mind. Historically, it has been prescribed for fatigue, insomnia, and respiratory ailments, but its most notable application lies in liver protection. It has long been used to treat liver disorders, including hepatitis and toxin-induced liver injury, aligning with modern pharmacological evidence demonstrating its potent hepatoprotective effects.

Aim of the study: This study aimed to develop a novel comprehensive two-dimensional primary Hepatic stellate cell (HSC) membrane chromatographic system to screen anti-fibrotic components from S. chinensis and to investigate the anti-fibrotic effects and mechanisms of gomisin N, a key lignan compound.

Materials and methods: A novel bioaffinity-based chromatographic screening system was designed and integrated with mass spectrometry to identify compounds with high HSC membrane affinity. The anti-fibrotic effects of the identified compound were then evaluated in vitro using primary HSCs and in vivo using a carbon tetrachloride-induced liver fibrosis mouse model. Primary HSC membrane chromatography coupled with high-resolution time-of-flight mass spectrometry was used to screen S. chinensis components. The affinity of the identified compounds was validated, and gomisin N was selected for further study. Cellular assays assessed its effects on HSC activation, while in vivo studies evaluated its impact on liver fibrosis. Mechanistic studies involved Western blotting, immunofluorescence, and real-time quantitative reverse transcription PCR to assess inflammation, oxidative stress, hepatic microvascular formation, and apoptosis-related pathways.

Results: Gomisin N exhibited the highest retention time on the HSC membrane chromatographic column, indicating strong HSC binding affinity. It significantly inhibited HSC activation and proliferation in vitro. In the CCl₄-induced liver fibrosis mouse model, gomisin N ameliorated liver fibrosis, reduced inflammatory cytokine expression, decreased oxidative stress markers, and suppressed hepatic microvascular formation. Mechanistically, it inhibited the Nuclear factor kappa B pathway, reduced hepatocyte apoptosis via regulation of B-cell lymphoma 2/Bcl-2-associated X protein and Caspase-3, and demonstrated a favourable safety profile.

Conclusion: This study presents a novel two-dimensional primary HSC membrane chromatographic system for screening anti-fibrotic compounds and is the first to report the anti-fibrotic effects of gomisin N. The findings provide experimental evidence supporting gomisin N as a promising lead compound for anti-fibrotic drug development.

Keywords: Cell membrane chromatography; Gomisin N; Hepatic stellate cells; Liver fibrosis; Schisandra chinensis (Turcz.) Baill.