College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
College of Medical, Nanjing University of Chinese Medicine, Nanjing, 210023, China; The First Clinical School of Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Phytomedicine. 2024 Dec;135:156098. doi: 10.1016/j.phymed.2024.156098. Epub 2024 Sep 28.
Hepatic fibrosis, a chronic pathological condition resulting from various forms of persistent liver injury, in the later stage, it can evolve into cirrhosis and even liver cancer. Curcumae Rhizoma (CR), traditionally recognized for its properties in line qi break blood, eliminate accumulation and relieve pain. According to traditional Chinese medicine (TCM) principles, vinegar-processing enhances CR's ability to enter the liver meridian and act on the blood level, potentially augmenting its therapeutic effects on hepatic diseases. Therefore, vinegar-processed Curcumae Rhizoma (VCR) is frequently employed in treating liver fibrosis and related hepatic conditions. However, the underlying mechanisms of vinegar processing in enhancing its therapeutic efficacy remain unclear.
The anti-liver fibrosis effects of CR and VCR were verified at individual and cellular levels. Subsequently, HPLC-Q-TOFMS and pharmacokinetic analysis were utilized to elucidate the potential bioactive substances underlying the enhanced anti-fibrotic efficacy of VCR. Building upon these findings, network pharmacology and metabolomics were integrated to screen for key effect components and regulatory pathways. Finally, the mechanisms of action were further analyzed and validated at the tissue and cellular levels through Western blotting (WB) and molecular docking studies.
Both CR and VCR exhibited therapeutic effects against hepatic fibrosis, with VCR demonstrating enhanced efficacy after vinegar processing. 6 sesquiterpenes including furanodiene and curdione, showed significant alterations in plasma exposure and hepatic distribution post-processing. VCR significantly improved pathological liver conditions, lipid accumulation, and fibrosis severity. Additionally, VCR markedly reduced the expression of α-SMA in the liver and attenuated the elevations in liver function markers such as ALT and AST. Combined network pharmacology, metabolomics, and hepatic tissue WB analysis revealed that the reduced phosphorylation of the PI3K/Akt/mTOR pathway is a critical mechanism in VCR's anti-fibrotic effects. Experiments on LX-2 cells demonstrated that four sesquiterpenes, including furanodiene and curdione, effectively inhibited the proliferation of activated hepatic stellate cells (HSCs). Furanodiene, in particular, promoted apoptosis in activated HSCs by reducing phosphorylation levels of the PI3K/Akt/mTOR pathway proteins, increasing BAX expression, and activating downstream caspase-3 to achieve the effect of anti-liver fibrosis.
Vinegar-processing significantly increases the plasma exposure and hepatic distribution of components such as furanodiene in VCR, enhancing anti-fibrotic efficacy by downregulating the phosphorylation levels of the PI3K/Akt/mTOR pathway and promoting HSC apoptosis. This study provides a comprehensive explanation of the vinegar-processing mechanism and its role in enhancing the anti-fibrotic effects of VCR, offering insights for its clinical application in liver fibrosis treatment and reference for the mechanistic study of other vinegar-processed herbal medicines.
肝纤维化是一种由各种形式的持续性肝损伤引起的慢性病理状态，在后期，它可能发展为肝硬化甚至肝癌。莪术（CR）传统上被认为具有行气破血、消积止痛的功效。根据中医（TCM）原理，醋制可增强 CR 入肝经、行血的作用，可能增强其治疗肝病的疗效。因此，醋制莪术（VCR）常被用于治疗肝纤维化及相关肝病。然而，醋制增强其疗效的潜在机制尚不清楚。
在个体和细胞水平上验证了 CR 和 VCR 的抗肝纤维化作用。随后，利用 HPLC-Q-TOFMS 和药代动力学分析阐明了 VCR 增强抗纤维化疗效的潜在生物活性物质。在此基础上，通过网络药理学和代谢组学筛选关键效应成分和调控通路。最后，通过 Western blot（WB）和分子对接研究在组织和细胞水平上进一步分析和验证作用机制。
CR 和 VCR 均对肝纤维化有治疗作用，VCR 经醋制后疗效增强。6 种倍半萜类化合物，包括呋喃二烯和莪术二酮，经醋制后血浆暴露和肝分布有明显变化。VCR 显著改善肝脏病理状况、脂质堆积和纤维化程度。此外，VCR 显著降低肝脏中 α-SMA 的表达，并降低 ALT 和 AST 等肝功能标志物的升高。联合网络药理学、代谢组学和肝组织 WB 分析表明，降低 PI3K/Akt/mTOR 通路的磷酸化是 VCR 抗纤维化作用的关键机制。LX-2 细胞实验表明，包括呋喃二烯和莪术二酮在内的四种倍半萜类化合物可有效抑制活化的肝星状细胞（HSCs）的增殖。特别是呋喃二烯通过降低 PI3K/Akt/mTOR 通路蛋白的磷酸化水平，增加 BAX 表达，激活下游 caspase-3，促进活化 HSCs 的凋亡，达到抗肝纤维化的效果。
醋制显著增加了 VCR 中呋喃二烯等成分的血浆暴露和肝分布，通过下调 PI3K/Akt/mTOR 通路的磷酸化水平和促进 HSC 凋亡，增强了抗纤维化作用。该研究全面解释了醋制机制及其在增强 VCR 抗纤维化作用中的作用，为其在肝纤维化治疗中的临床应用提供了依据，并为其他醋制草药的机制研究提供了参考。
