Bioactive fraction of processed Curcumae Rhizoma-Sparganii Rhizoma anti-liver fibrosis by regulate taurine metabolism through PI3K/AKT pathway

Abstract

Ethnopharmacological relevance: Fibrosis is a prevalent pathological process in various chronic liver conditions. Curcumae Rhizoma-Sparganii Rhizoma (CR-SR) and its vinegar-processed derivatives are representative traditional Chinese medicines (TCM) that are utilized for promoting blood circulation, eliminating stasis and anti liver fibrosis in clinical practice in China. However, the bioactive fraction and the action of mechanism of CR-SR remain elusive.

Aim of study: This study aims to systemic compare the pharmacological action of single herb, herb pairs, and vinegar-processed herb pairs of CR-SR. Afterwards, we intend to screen out the anti-liver fibrosis bioactive fraction of vinegar processed CR-SR (VP-CRSR) and preliminarily clarify the bioactive mechanism.

Materials and methods: The anti-liver fibrosis effect of CR, SR, CR-SR, VP-CRSR and different polar fractions of VP-CRSR were compared by a CCL₄-induced mouse model. The change of material basis was analyzed by UPLC. The chemical composition of optimum active fraction was identified by UPLC-Q-TOF-MS combined with GNPS. Plasma metabolomics were conducted to explore the potential active mechanism of the optimum effective fraction of VP-CRSR. Molecular biological techniques were employed to validate the anti-hepatic fibrosis molecular mechanisms of the optimal effective fraction both in vivo and in vitro.

Results: The pharmacological study results showed that VP-CRSR has a better anti-fibrosis effect compared to each single herb and crude herb pair. Further liver fibrosis mice experiment illustrates that ethyl acetate extract of VP-CRSR (VEA) turned out to be the optimum anti-fibrosis bioactive fraction which can achieve curative effect as VP-CRSR. UPLC fingerprint exhibits great changes before and after vinegar processed of CR, SR, and CR-SR. UPLC-Q-TOF-MS combined with GNPS analysis results indicate that the main active ingredients of VEA mainly include procurcumadiol, procurcumenol, zedoarondiol, formononetin etc. Metabolomics study indicated that VEA optimum active fractions could significantly regulate taurine and hypotaurine metabolism to ameliorate liver fibrosis. Further molecular mechanism study in vivo and in vitro shows that PI3K/AKT/eNOS and TGF-β1/Smad signaling pathways have important value in taurine and hypotaurine metabolism which regulated by VEA.

Conclusion: This study systematically elucidates the material foundation and potential mechanisms of action of VP-CRSR in combating liver fibrosis from a holistic perspective. The research findings offer robust theoretical support for the development and utilization of the CR-SR herbal pair. Furthermore, this study introduces a valuable research methodology for investigating the active components and bioactive mechanisms of TCM.

Keywords: Bioactive fraction; Curcumae Rhizoma-Sparganii Rhizoma; Liver fibrosis; PI3K/AKT/eNOS signal pathway; TGF-β1/Smad signal pathway; Taurine and hypotaurine metabolism.