Gao Siqi, Wang Xingxing, Xu Qiuying, Li Rongsheng, Yao Lumeng, Zhang Anna, Zhou Qun, Xiao Zhun, Li Shengsheng, Meng Xiongyu, Wu Jianjun, Qin Luping
School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 311402, China.
Department of Digestive Diseases, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, China.
J Ethnopharmacol. 2025 Apr 9;345:119571. doi: 10.1016/j.jep.2025.119571. Epub 2025 Feb 27.
Sanghuangporus, the dried fruiting body of Sanghuangporus vaninii (Ljub) L.W.Zhou et Y.C.Dai. As the main species of Sanghuang, it has been well-known and used commonly as a traditional medicinal and edible macrofungi for thousands of years in many countries, including China, Korea and Japan. Although it has good hepatoprotective activity, its potential efficacy and mechanism on liver fibrosis remain elusive.
Total Sanghuangporus vaninii extract (TSH) was prepared by ethanol extraction to investigate its chemical components and to conduct an initial assessment of its efficacy and underlying mechanism in a murine model of liver fibrosis.
The chemical components of TSH were initially analyzed by UHPLC-Q-Orbitrap HRMS. To elucidate the effects of TSH, an in vivo model of fibrosis was established in mice using carbon tetrachloride (CCl), followed by assessments of serum liver function and histopathological analysis. Besides, indicators related to liver fibrosis, hepatic stellate cells (HSCs) activation, inflammation response and ferroptosis related indicators were detected by western blotting, immunohistochemistry and real-time quantitative PCR (RT-qPCR) analysis. Additionally, the 16S rDNA sequencing and untargeted metabolomics analysis of intestinal microbiota were employed to investigating the role of TSH in gut microbiome. In vitro, the human hepatocyte line L02 was stimulated with erastin and treated with or without TSH to elucidate its underlying mechanism.
The administration of TSH significantly improved serum indicators of liver injury in CCl-induced fibrosis mice, reduced HSCs activation and collagen deposition, while inhibiting the expressions of transforming growth factor-β1(TGF-β1)/Smad signaling pathway. Notably, TSH treatment attenuated hepatocyte ferroptosis and lipid peroxidation both in vivo and in vitro, as evidenced by a marked decrease in liver iron and malondialdehyde (MDA) contents. In particular, TSH was demonstrated to activate the nuclear factor erythroid 2-related factor 2 (Nrf2)-glutathione peroxidase 4 (GPX4) signaling pathway, thereby protecting hepatocytes from ferroptosis with a particular enhancement of Nrf2 nuclear transcription. Furthermore, TSH influenced gut microbiota composition and ameliorated intestinal metabolic disorders. The increased abundance of Parasutterella and Olsenellas due to TSH treatment was significantly positively correlated with elevated phosphatidylcholines involved in linoleic acid metabolism, and negatively correlated with the reduction of fatty acyls. And the enrichment of intestinal linoleic acid metabolism presented a negative correlation in liver fibrosis biomarkers.
Our findings indicate that the TSH treatment exerts a significantly protective effect on CCl-induced mice by ameliorating hepatic injury and ferroptosis damage, inhibiting HSCs activation and collagen deposition, and remodeling gut microbiota homeostasis and metabolic imbalance. Notably, TSH attenuated hepatocyte ferroptosis in liver fibrosis and exhibited upregulation of the Nrf2-GPX4 signaling pathway. Furthermore, TSH could enrich the abundance of Parasutterella and Olsenellas, which may contribute to intestinal linoleic acid metabolism, thereby contributing to the reduction of liver fibrosis damage. Our study provides more effective and unreported evidence of TSH in anti-fibrosis activity.
桑黄是瓦尼桑黄（Ljub）L.W.周和Y.C.戴的干燥子实体。作为桑黄的主要种类，它在包括中国、韩国和日本在内的许多国家已经闻名并作为传统药食两用大型真菌被广泛使用了数千年。尽管它具有良好的肝脏保护活性，但其对肝纤维化的潜在功效和机制仍不清楚。
通过乙醇提取制备总瓦尼桑黄提取物（TSH），以研究其化学成分，并在小鼠肝纤维化模型中对其功效及潜在机制进行初步评估。
首先通过超高效液相色谱-四极杆-轨道阱高分辨质谱（UHPLC-Q-Orbitrap HRMS）分析TSH的化学成分。为阐明TSH的作用，使用四氯化碳（CCl）在小鼠中建立纤维化体内模型，随后评估血清肝功能并进行组织病理学分析。此外，通过蛋白质免疫印迹法、免疫组织化学和实时定量聚合酶链反应（RT-qPCR）分析检测与肝纤维化、肝星状细胞（HSCs）活化、炎症反应和铁死亡相关的指标。另外，采用16S核糖体脱氧核糖核酸（rDNA）测序和肠道微生物群的非靶向代谢组学分析来研究TSH在肠道微生物群中的作用。在体外，用人肝细胞系L02用埃拉斯汀刺激并分别用或不用TSH处理，以阐明其潜在机制。
给予TSH可显著改善CCl诱导的纤维化小鼠的肝损伤血清指标，减少HSCs活化和胶原沉积，同时抑制转化生长因子-β1（TGF-β1）/Smad信号通路的表达。值得注意的是，TSH处理在体内和体外均减轻了肝细胞铁死亡和脂质过氧化，肝脏铁和丙二醛（MDA）含量显著降低证明了这一点。特别是，TSH被证明可激活核因子红细胞2相关因子2（Nrf2）-谷胱甘肽过氧化物酶4（GPX4）信号通路，从而通过特别增强Nrf2核转录来保护肝细胞免受铁死亡。此外，TSH影响肠道微生物群组成并改善肠道代谢紊乱。TSH处理导致的副萨特氏菌属和奥尔森氏菌属丰度增加与参与亚油酸代谢的磷脂酰胆碱升高显著正相关，与脂肪酰基的减少负相关。并且肠道亚油酸代谢的富集与肝纤维化生物标志物呈负相关。
我们的研究结果表明，TSH处理通过改善肝损伤和铁死亡损伤、抑制HSCs活化和胶原沉积以及重塑肠道微生物群稳态和代谢失衡，对CCl诱导的小鼠具有显著的保护作用。值得注意的是，TSH减轻了肝纤维化中的肝细胞铁死亡，并表现出Nrf2-GPX4信号通路的上调。此外，TSH可增加副萨特氏菌属和奥尔森氏菌属的丰度，这可能有助于肠道亚油酸代谢，从而有助于减少肝纤维化损伤。我们的研究为TSH的抗纤维化活性提供了更有效且未报道的证据。
