Pharmacology Laboratory of Prevention and Treatment of High Incidence of Disease, Guilin Medical University, Guilin, 541199, Guangxi, China.
Pharmacology Laboratory of Prevention and Treatment of High Incidence of Disease, Guilin Medical University, Guilin, 541199, Guangxi, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 3):118972. doi: 10.1016/j.jep.2024.118972. Epub 2024 Oct 23.
Cholestatic liver disease (CLD) is a disease characterized by cholestasis. Farnesoid X receptor (FXR) is a nuclear receptor that maintains homeostasis in bile acid metabolism. Studies have shown that gut microbiota interfered with the FXR pathway. Modulation of FXR to inhibit cholestasis has become a key measure in the treatment of CLD. In traditional folk medicine, Desmodium styracifolium (Osb.) Merr. was used as a primary treatment for gallstones, gonorrhea, jaundice, cholecystitis and other diseases. Modern pharmacological studies had also found that the herb has anti-calculus, anti-inflammatory, antioxidant, diuretic and liver damage. Therefore, we speculated that Desmodium styracifolium (Osb.) Merr. extracts (DME) could alleviate CLD through the FXR pathway and might be associated with the gut microbiota. However, studies of DME alleviating CLD through the FXR pathway have not been reported.
To study the effect and mechanism of DME in relieving CLD through in vivo and in vitro experiments.
First, mice were administrated with alpha-naphthyl isothiocyanate (ANIT) to establish a CLD model in vivo. Meanwhile, HepG2 cells were induced by lithocholic acid (LCA) to establish the CLD model in vitro. To evaluate the therapeutic effect of DME on CLD mice, hematoxylin-eosin (HE) staining, and biochemical indicators were performed. The prototype of the blood components in mice serum was detected by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). 16S rDNA sequencing was used to analyze the gut microbiota. Finally, the protein and mRNA expression of the FXR pathway in mice liver tissues or HepG2 cells were detected by Western blot, qRT-PCR, or immunofluorescence.
Pathological testing and biochemical indexes showed that DME significantly reduced serum ALT, AST, ALP, TBIL, DBIL, TBA and liver TBA levels, and attenuated liver tissue injury, necrosis and jaundice in CLD mice. In addition, MetagenomeSeq analysis of gut microbiota showed that DME significantly up-regulated the abundance of Parvibacter, down-regulated the abundance of Paenalcaligenes, and regulated bile acid homeostasis. In terms of mRNA expression, DME significantly upregulated the mRNA levels of Nr1h4, Abcb11, Cyp7a1 and Slc10a1. Meanwhile, in terms of protein expression, DME significantly up-regulated the protein expression levels of FXR, BSEP, CYP7A1 and NTCP, which regulated bile acid homeostasis. Finally, the molecular docking results showed that the components of DME, such as Lumichrome, Daidzein and Folic acid, all had good binding ability with FXR, and the surface plasmon resonance (SPR) results also showed that both Lumichrome and Daidzein had a relatively high affinity with FXR.
DME alleviated CLD through the FXR pathway, and the mechanisms might be associated with the gut microbiota.
ETHNOPHARMACOLOGICAL 相关性：胆汁淤积性肝病 (CLD) 是一种以胆汁淤积为特征的疾病。法尼醇 X 受体 (FXR) 是一种核受体，可维持胆汁酸代谢的内稳态。研究表明，肠道微生物群会干扰 FXR 途径。抑制胆汁淤积的 FXR 调节已成为治疗 CLD 的关键措施。在传统民间医学中，野葛（Osb.） Merr. 被用作胆结石、淋病、黄疸、胆囊炎等疾病的主要治疗方法。现代药理学研究还发现，该草药具有抗结石、抗炎、抗氧化、利尿和肝损伤作用。因此，我们推测野葛（Osb.） Merr. 提取物（DME）可能通过 FXR 途径缓解 CLD，并且可能与肠道微生物群有关。然而，尚未有研究报道 DME 通过 FXR 途径缓解 CLD。
通过体内和体外实验研究 DME 缓解 CLD 的作用和机制。
首先，用α-萘基异硫氰酸酯 (ANIT) 在体内建立 CLD 模型。同时，用胆酸 (LCA) 诱导 HepG2 细胞在体外建立 CLD 模型。为了评估 DME 对 CLD 小鼠的治疗效果，进行了苏木精-伊红 (HE) 染色和生化指标检测。采用超高效液相色谱-串联质谱 (UPLC-MS/MS) 检测小鼠血清中原型血成分。采用 16S rDNA 测序分析肠道微生物群。最后，通过 Western blot、qRT-PCR 或免疫荧光检测小鼠肝组织或 HepG2 细胞中 FXR 通路的蛋白和 mRNA 表达。
病理检查和生化指标表明，DME 可显著降低 CLD 小鼠血清 ALT、AST、ALP、TBIL、DBIL、TBA 和肝 TBA 水平，减轻肝组织损伤、坏死和黄疸。此外，肠道微生物群的 MetagenomeSeq 分析表明，DME 可显著上调 Parvibacter 的丰度，下调 Paenalcaligenes 的丰度，并调节胆汁酸稳态。在 mRNA 表达方面，DME 可显著上调 Nr1h4、Abcb11、Cyp7a1 和 Slc10a1 的 mRNA 水平。同时，在蛋白表达方面，DME 可显著上调 FXR、BSEP、CYP7A1 和 NTCP 的蛋白表达水平，从而调节胆汁酸稳态。最后，分子对接结果表明，DME 的成分，如 Lumichrome、大豆苷元和叶酸，均与 FXR 具有良好的结合能力，表面等离子体共振 (SPR) 结果也表明，Lumichrome 和大豆苷元与 FXR 均具有较高的亲和力。
DME 通过 FXR 途径缓解 CLD，其机制可能与肠道微生物群有关。
