目的 基于肝星状细胞（hepatic stellate cells，HSCs）铁代谢研究沙苑子总黄酮（total flavonoids from Astragali Complanati Semen，TFACS）对肝纤维化（hepatic fibrosis，HF）小鼠的影响。方法 小鼠连续8周ip 10%四氯化碳（carbon tetrachloride，CCl₄）建立HF小鼠模型，HF小鼠随机分为对照组、模型组、水飞蓟宾（100 mg/kg）组和TFACS低、高剂量（8、20 mg/kg）组，连续给药6周，末次给药16 h后收集血清，计算肝脏和脾脏指数；采用全自动血清生化仪测定血清肝功能指标；采用ELISA试剂盒检测小鼠血清中Ⅳ型胶原蛋白（collagen type IV，Col-IV）、透明质酸（hyaluronic acid，HA）、层黏蛋白（laminin，LN）和Ⅲ型前胶原（procollagen Ⅲ，PCⅢ）水平；采用苏木素-伊红（HE）和Masson染色观察肝组织病变和胶原沉积；采用试剂盒测定肝组织中铁调素、铁以及丙二醛（malondialdehyde，MDA）含量；采用普鲁士蓝染色观察肝组织中铁的分布；采用Western blotting检测肝组织中Ⅰ型胶原蛋白（CollagenⅠ）、Ⅳ型胶原蛋白（Collagen Ⅳ）、α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）和转铁蛋白受体1（transferrin receptor 1，TFRC）的表达。采用转化生长因子-β1（transforming growth factor-β1，TGF-β1）诱导HSC-T6细胞活化，CCK-8法检测TFACS对细胞增殖的影响，试剂盒测定细胞中MDA和铁含量，采用免疫荧光法检测细胞中活性氧（reactive oxygen species，ROS）水平，采用Western blotting检测细胞中α-SMA和TFRC蛋白的表达。结果 TFACS显著降低HF小鼠的脾脏指数（P＜0.05、0.01），降低血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase，AST）、碱性磷酸酶（alkaline phosphatase，ALP）活性和Col-Ⅳ、HA、LN、PCⅢ水平（P＜0.05、0.01、0.001），改善HF小鼠肝细胞损伤、炎症浸润和胶原沉积，减少肝组织铁染色面积（P＜0.05），降低肝组织中铁调素、铁和MDA水平（P＜0.05、0.01），并下调肝组织Collagen Ⅰ、Collagen Ⅳ、TFRC和α-SMA蛋白表达（P＜0.05）。此外，TFACS可明显抑制TGF-β1诱导的HSC-T6细胞增殖（P＜0.05、0.01），下调TFRC和α-SMA蛋白表达（P＜0.05、0.01），并降低MDA、ROS和铁水平（P＜0.001）。结论 TFACS对HF具有保护作用，其作用机制可能与调节HSCs铁代谢有关。

Objective To study the effect of total flavonoids from Shayuanzi (Astragali Complanati Semen, TFACS) on hepatic fibrosis (HF) mice based on iron metabolism in hepatic stellate cells (HSCs). Methods HF mice model was established by ip 10% carbon tetrachloride (CCl₄) for eight consecutive weeks. HF mice were randomly divided into control group, model group, silibinin (100 mg/kg) group, and TFACS low-, high-dose (8, 20 mg/kg) groups. The mice were administered continuously for six weeks, and serum was collected 16 h after the last administration. Liver and spleen indexes were calculated; Serum liver function index was measured by automatic serum biochemical instrument; ELISA was used to measure the levels of collagen type IV (Col-IV), hyaluronic acid (HA), laminin (LN) and procollagen Ⅲ (PCⅢ) in serum; The liver lesions and collagen deposition were observed by hematoxylin-eosin (HE) and Masson staining; The contents of hepcidin, iron and MDA in liver tissue were determined by kit; The distribution of iron in liver tissue was observed by prussian blue staining; Western blotting was used to detect the expressions of Collagen Ⅰ, Collagen Ⅳ, α-smooth muscle actin (α-SMA) and transferrin receptor (TFRC) in liver tissue. Transforming growth factor-β1 (TGF-β1) was used to induce the activation of HSC-T6 cells, and the effect of TFACS on the proliferation of cells was evaluated by CCK-8 method, the contents of MDA and iron in cells were determined by kits, the level of reactive oxygen species (ROS) in cells was detected by immunofluorescence method, the expressions of α-SMA and TFRC proteins in cells were detected by Western blotting. Results TFACS significantly reduced the spleen index of HF mice (P < 0.05, 0.01), reduced the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and levels of Col-IV, HA, LN and PCIII in serum (P < 0.05, 0.01, 0.001), improved liver cell injury, inflammatory infiltration and collagen deposition in HF mice, reduced the iron staining area of liver tissue (P < 0.05), decreased the levels of hepcidin, iron and MDA in liver tissue (P < 0.05, 0.01), and down-regulate the expressions of Collagen I, Collagen IV, TFRC and α-SMA proteins in liver tissue (P < 0.05). In addition, TFACS could significantly inhibit the proliferation of HSC-T6 cells induced by TGF-β1 (P < 0.05, 0.01), down-regulate the expressions of TFRC and α-SMA proteins (P < 0.05, 0.01), and reduce MDA, ROS and iron levels (P < 0.001). Conclusion TFACS has a protective effect on HF, and its mechanism may be related to regulating iron metabolism in HSCs.

R285.5

陕西省科技厅重点研发计划项目（2024SF-YBXM-458，2024NC-YBXM-263）；陕西省教育厅重点科学研究计划项目（22JY019）；陕西省科技创新团队（2022TD-56）；陕西高校青年创新团队建设项目（2023）；秦创原中医药产业创新聚集区项目（L2024-QCY-ZYYJJQ-X201）；陕西省重点研发计划-重点产业创新链（群）（社会发展领域）（2023-ZDLSF-58）