Hao Min, Chen Ziyan, Wang Pingping, Lu Yandan, An Dongyang, Yang Qiao, Han Xin, Wang Kuilong, Sang Xianan, Cao Gang
School of Pharmaceutical Sciences, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310053, China.
School of Pharmaceutical Sciences, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310053, China.
J Ethnopharmacol. 2025 Jul 24;351:120090. doi: 10.1016/j.jep.2025.120090. Epub 2025 Jun 3.
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.
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.
The anti-liver fibrosis effect of CR, SR, CR-SR, VP-CRSR and different polar fractions of VP-CRSR were compared by a CCL-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.
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.
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.
纤维化是各种慢性肝脏疾病中普遍存在的病理过程。莪术-三棱及其醋制衍生物是中国临床实践中用于促进血液循环、化瘀和抗肝纤维化的代表性传统中药。然而，莪术-三棱的生物活性成分及其作用机制仍不清楚。
本研究旨在系统比较莪术-三棱单味药、药对及醋制后药对的药理作用。之后，我们打算筛选出醋制莪术-三棱（VP-CRSR）的抗肝纤维化生物活性成分，并初步阐明其生物活性机制。
通过CCL诱导的小鼠模型比较莪术（CR）、三棱（SR）、莪术-三棱（CR-SR）、VP-CRSR及VP-CRSR不同极性部位的抗肝纤维化作用。采用超高效液相色谱法（UPLC）分析物质基础的变化。通过UPLC-Q-TOF-MS结合全球天然产物社会分子网络（GNPS）鉴定最佳活性部位的化学成分。进行血浆代谢组学研究以探索VP-CRSR最佳有效部位的潜在活性机制。采用分子生物学技术在体内和体外验证最佳有效部位抗肝纤维化的分子机制。
药理研究结果表明，与各单味药和生药对相比，VP-CRSR具有更好的抗纤维化作用。进一步的肝纤维化小鼠实验表明，VP-CRSR的乙酸乙酯提取物（VEA）是最佳的抗纤维化生物活性成分，其疗效与VP-CRSR相当。UPLC指纹图谱显示CR、SR和CR-SR醋制前后有很大变化。UPLC-Q-TOF-MS结合GNPS分析结果表明，VEA的主要活性成分主要包括莪术二酮、莪术烯醇、泽兰二醇、芒柄花素等。代谢组学研究表明，VEA最佳活性部位可显著调节牛磺酸和低牛磺酸代谢以改善肝纤维化。进一步的体内和体外分子机制研究表明，PI3K/AKT/eNOS和TGF-β1/Smad信号通路在VEA调节的牛磺酸和低牛磺酸代谢中具有重要作用。
本研究从整体角度系统阐明了VP-CRSR抗肝纤维化的物质基础和潜在作用机制。研究结果为莪术-三棱药对的开发利用提供了有力的理论支持。此外，本研究为研究中药活性成分和生物活性机制引入了一种有价值的研究方法。
