Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Institute of Liver Diseases, Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai, 201203, China; Department of Pharmacy, The SATCM Third Grade Laboratory of Traditional Chinese Medicine Preparations, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai, 201203, China.
Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Institute of Liver Diseases, Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai, 201203, China.
J Ethnopharmacol. 2024 Dec 5;335:118649. doi: 10.1016/j.jep.2024.118649. Epub 2024 Jul 31.
Cholestatic liver diseases (CLD) are liver disorders resulting from abnormal bile formation, secretion, and excretion from various causes. Due to the lack of suitable and safe medications, liver transplantation is the ultimate treatment for CLD patients. Isoastragaloside I (IAS I) is one of the main saponin found in Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge, which has been demonstrated to obviously alleviate CLD. Nevertheless, the IAS I's specific anti-CLD mechanism remains undecipherable.
This study's purpose was to elucidate the protective consequence of IAS I on 0.1% 3, 5-diethoxycarbonyl-1,4-dihydroxychollidine (DDC) diet-induced CLD mice, and to reveal its potential mechanism.
In this study, mice with CLD that had been fed a 0.1% DDC diet were distributed two doses of IAS I (20 mg/kg, 50 mg/kg). The effects of IAS I on CLD models were investigated by assessing blood biochemistry, liver histology, and Hyp concentrations. We investigated markers of liver fibrosis and ductular reaction using immunohistochemistry, Western blot, and qRT-PCR. Liver inflammation indicators, arachidonic acid (ARA), and ω-3 fatty acid (FA) metabolites were also analyzed. Quantitative determination of 39 bile acids (BAs) in different organs employing UHPLC-Q-Exactive Orbitrap HRMS technology. Additionally, the H&E and Western blot analysis were used to evaluate differences in intestinal barrier function in DDC-induced mice before and after administering IAS I.
After treatment with IAS I, serum biochemical indicators and liver hydroxyproline (Hyp) increased in a dose-dependent manner in CLD mice. The IAS I group showed significant improvement in indicators of liver fibrosis and ductular response, including as α-smooth muscle actin (α-SMA) and cytokeratin 19 (CK19), and transforming growth factor-β (TGF-β)/Smads signaling pathway. And inflammatory factors: F4/80, tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), ARA and ω-3 FA metabolites showed significant improvement following IAS I treatment. Moreover, IAS I significantly ameliorated liver tau-BAs levels, particularly TCA, THCA, THDCA, TCDCA, and TDCA contents, which were associated with enhanced expression of hepatic farnesoid X receptor (FXR), small heterodimer partner (SHP), cholesterol 7α-hydroxylase (Cyp7a1), and bile-salt export pump (BSEP). Furthermore, IAS I significantly improved pathological changes and protein expression related to intestinal barrier function, including zonula occludens protein 1 (ZO-1), Muc2, and Occludin.
IAS I alleviated cholestatic liver injury, relieved inflammation, improved the altered tau-BAs metabolism and restored intestinal barrier function to protect against DDC-induced cholestatic liver diseases.
ETHNOPHARMACOLOGICAL 相关性：胆汁淤积性肝病（CLD）是由各种原因导致胆汁形成、分泌和排泄异常引起的肝脏疾病。由于缺乏合适和安全的药物，肝移植是 CLD 患者的最终治疗方法。异黄芪甲苷 I（IAS I）是黄芪（Fisch.）Bge. var.蒙古（Bge.）Hsiao 或黄芪（Fisch.）Bge 中发现的主要皂苷之一，已被证明可明显缓解 CLD。然而，IAS I 的具体抗 CLD 机制仍未被破译。
本研究旨在阐明 IAS I 对 0.1%3,5-二乙氧羰基-1,4-二羟基胆甾烷（DDC）饮食诱导的 CLD 小鼠的保护作用，并揭示其潜在机制。
在这项研究中，用 0.1%DDC 饮食喂养的 CLD 小鼠被分为两个剂量的 IAS I（20mg/kg，50mg/kg）。通过评估血液生化、肝组织学和 Hyp 浓度来研究 IAS I 对 CLD 模型的影响。我们使用免疫组织化学、Western blot 和 qRT-PCR 研究了肝纤维化和胆管反应的标志物。还分析了肝炎症指标、花生四烯酸（ARA）和 ω-3 脂肪酸（FA）代谢物。采用 UHPLC-Q-Exactive Orbitrap HRMS 技术定量测定不同器官中的 39 种胆汁酸（BA）。此外，还使用 H&E 和 Western blot 分析评估了 DDC 诱导的小鼠在给予 IAS I 前后肠道屏障功能的差异。
在 CLD 小鼠中，IAS I 治疗后，血清生化指标和肝脏羟脯氨酸（Hyp）呈剂量依赖性增加。IAS I 组肝纤维化和胆管反应的标志物，包括α-平滑肌肌动蛋白（α-SMA）和细胞角蛋白 19（CK19）以及转化生长因子-β（TGF-β）/Smads 信号通路均有显著改善。炎症因子：F4/80、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、ARA 和 ω-3 FA 代谢物在 IAS I 治疗后均有显著改善。此外，IAS I 显著改善了肝脏 tau-BAs 水平，特别是 TCA、THCA、THDCA、TCDCA 和 TDCA 含量，这与肝脏法尼醇 X 受体（FXR）、小异二聚体伴侣（SHP）、胆固醇 7α-羟化酶（Cyp7a1）和胆汁盐输出泵（BSEP）的表达增强有关。此外，IAS I 还显著改善了与肠道屏障功能相关的病理变化和蛋白表达，包括紧密连接蛋白 1（ZO-1）、Muc2 和 Occludin。
IAS I 缓解了胆汁淤积性肝损伤，减轻了炎症，改善了异常的 tau-BAs 代谢，并恢复了肠道屏障功能，从而预防了 DDC 诱导的胆汁淤积性肝病。
Zotero 插件