目的 探究一贯煎多糖（YGJP -W）的基本结构特征以及其改善四氯化碳（CCl₄）诱导小鼠肝纤维化的活性，分析YGJP-W抗肝纤维化活性与其调节肠道菌群结构变化关系。方法 采用水提醇沉和弱阴离子交换色谱制备一贯煎多糖组分YGJP-W，依次采用苯酚-硫酸法、间羟基联苯法、BCA蛋白定量分析试剂盒、高效凝胶渗透色谱法（high performance gel permeation chromatography，HPGPC）、FT-IR、离子色谱仪及扫描电镜法（scanning electron microscope，SEM）对一贯煎多糖的总糖含量、糖醛酸含量、蛋白含量、相对分子质量分布、官能团、单糖组成及形貌特征进行表征，采用CCl₄诱导的小鼠模型，检测YGJP-W（50、100 mg/kg）干预3周后的小鼠血清谷丙转氨酶、谷草转氨酶及肝组织羟脯氨酸水平，并通过HE染色和天狼猩红染色观察肝脏组织学变化，评价YGJP-W改善肝纤维化活性；采用16S rDNA测序技术评价YGJP-W对小鼠肠道菌群的影响；采用抗生素联合YGJP-W干预，评价YGJP-W是否通过调节肠道菌群发挥药效作用。结果 YGJP-W总得率为11.60%，总糖质量分数为92.48%，至少由相对分子质量为1.74×10⁵和5.6×10³的2种多糖组成，单糖组成包含半乳糖、葡萄糖、甘露糖和果糖，其物质的量比为1.0∶2.8∶1.4∶1.6。YGJP-W干预3周后，能够显著改善CCl₄引起的肝功能异常、肝组织病理损伤、胶原沉积和纤维化。YGJP-W在改善肝纤维化的同时能够引起小鼠肠道菌群结构变化，在门水平上，YGJP-W显著性降低拟杆菌门丰度[（57.4±14.6）% vs（79.3±8.1）%，P＜0.01]，增加变形菌门[（10.4±4.7）% vs（2.1±1.9）%，P＜0.01] 和放线菌门[（3.7±2.4%）% vs（0.3±0.1）%，P＜0.01）]丰度，在属水平上，YGJP-W增加狄氏副拟杆菌属Parabacteroides、埃希氏菌-志贺氏菌属Escherichia-Shigella、活泼瘤胃球菌属Ruminococcus_gnavus_group、萨特氏菌属Sutterella、毛螺菌属NK4A136组Lachnospiraceae_NK4A136_group和梭杆菌属Fusobacterium共6个菌属的丰度（LDA score＞3，P＜0.01），降低拟普雷沃氏菌属Alloprevotella），肠鼠杆菌属Muribaculum，普雷沃氏菌属UCG-001 Prevotellaceae_UCG-001和另枝杆菌Alistipes共4个菌属的丰度（LDA score＞3，P＜0.01）。抗生素联合YJGP-W干预，YJGP-W抗肝纤维化的活性显著减弱。结论 多糖类成分YJGP-W是一贯煎抗肝纤维化的重要物质基础，YJGP-W发挥抗肝纤维化的作用可能与其调控肠道菌群相关。

Objective To evaluate the structural characterization of Yiguan Decoction polysaccharides (YGJP-W) and its effects on carbon tetrachloride (CCl₄)-induced liver fibrosis in mice, and to explore the relationship between the anti-liver fibrosis effect of YGJP-W and its regulation of gut microbiota structure. Methods YJGP-W was prepared by boiling water extraction, alcohol precipitation, and diethylaminoethyl-Sepharose Fast Flow anion exchange chromatography. The total sugar content, uronic acid content, protein content, molecular weight distribution, functional group composition, monosaccharide composition and morphological characteristics of YJGP-W were investigated by phenol-sulfuric acid method, m-hydroxy-biphenyl method, BCA protein quantitative analysis kit, high performance gel permeation chromatography (HPGPC), FT-IR, ion chromatograph and scanning electron microscope (SEM). The CCl₄-induced mouse model was used to detect the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in serum and hydroxyproline (Hyp) in liver tissue of mice after YJGP-W (50 mg/kg and 100 mg/kg) intervention for 3 weeks. Hematoxylin-eosin staining and sirius red staining were used to observe liver histological changes and to evaluate anti-liver fibrosis effect of YJGP-W. 16S rDNA sequencing technology was used to evaluate the effects of YJGP-W on gut microbiota in mice. And antibiotics combined with YJGP-W were used to evaluate whether YJGP-W exerted its medicinal effects by regulating gut microbiota. Results The total yield of YGJP-W was 11.60%, the total sugar content was 92.48%, and it was composed of at least two polysaccharides with molecular weights of 1.74 × 10⁵and 5.6 × 10³. Monosaccharide composition is composed of galactose, glucose, mannose, and fructose in molar ratios of 1.0∶2.8∶1.4∶1.6. YGJP-W could ameliorate CCl₄-induced liver dysfunction, liver tissue pathological damage, collagen deposition and fibrosis after intervention for three weeks. YGJP-W induced structural changes of gut microbiota in mice while ameliorateing liver fibrosis. At the phylum level, YGJP-W decreased the abundance of Bacteroidetes [ (57.4±14.6)% vs (79.3±8.1)%, P < 0.01] and increased the abundances of Proteobacteria [(10.4±4.7)% vs (2.1±1.9)%, P < 0.01] and Actinobacteria [(3.7±2.4)% vs (0.3±0.1)%, P < 0.01]. At the genus level, YGJP-W increased the abundances of six bacterial genera, including Parabacteroides,Escherichia-Shigella, Ruminococcus_gnavus_group, Sutterella,Lachnospiraceae_NK4A136_group and Fusobacterium (LDA score > 3,P < 0.01), and decreased the abundances of four bacterial genera, including Alloprevotella, Muribaculum, Prevotellaceae_UCG-001 and Alistipes (LDA score > 3, P < 0.01). After the intervention of antibiotics combined with YJGP-W, the anti-liver fibrosis activity of YJGP-W was weakened. Conclusion YJGP-W is an important material basis for anti-liver fibrosis effect of Yiguan Decoction, and the anti-liver fibrosis effect of YJGP-W may be related to its regulatory activity of gut microbiota.

国家自然科学基金项目（82130120）；国家自然科学基金项目（82004162）；国家自然科学基金项目（U23A20516）；上海市科技创新行动计划项目（20S21902600）；上海市复方中药重点实验室（上海中医药大学）开放课题基金（21DZ2270500）