目的 基于肠道微生物群、粪便代谢组学及网络药理学探讨藏药甲嘎松汤（干姜、豆蔻、肉豆蔻组成）对 酒精性肝病小鼠的作用及机制。方法 （1）采用酒精液体饲料灌胃 28 d 诱导酒精性肝病小鼠模型。将 C57BL/6 小鼠随机分为对照组、模型组、水飞蓟素组及甲嘎松汤低、中、高剂量组，每组 6 只。造模 1 周后，开始灌胃 给药，甲嘎松汤低、中、高剂量组分别给予 0.17、0.33、0.66 g·kg-1 甲嘎松汤水煎液灌胃，水飞蓟素组按 50 mg·kg-1灌胃，每日 1 次，连续给药 21 d。采用生化法检测肝脏组织甘油三酯（TG）、丙二醛（MDA）、低密度 脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）水平； 油红 O 染色法观察肝脏组织脂质累积情况；HE 染色法观察肝脏组织病理变化；进行肠道菌群 16S rRNA 基因 测序及粪便样品代谢谱分析。（2）采用网络药理学方法预测甲嘎松汤治疗酒精性肝病的潜在作用靶点，筛选出核 心活性成分、核心靶点。进行代谢组学结合网络药理学分析，构建“差异代谢物-反应-酶-基因”网络，筛选 色氨酸代谢的关键靶点，与网络药理学核心靶标取交集；对核心活性成分去氢二异丁香酚与核心靶点 CYP1A1、MAOA、MAOB 进行分子对接验证。（3）采用去氢二异丁香酚低、中、高剂量（6.25、12.5、25 µmol·L-1 ） 分别作用于 AML-12 细胞 24 h 后，用 400 mmol·L-1乙醇诱导 2 h。采用生化法检测细胞 TG、总胆固醇（TC）、 MDA、ALT、AST 水平；qRT-PCR 法检测细胞 CYP1A1、MAOA、MAOB mRNA 表达水平。结果 （1）与对照 组比较，模型组小鼠肝脏组织 TG、MDA、LDL-C、AST、ALT 水平均显著升高（P＜0.05，P＜0.01，P＜ 0.001），HDL-C 水平显著降低（P＜0.001）；肝脏组织中肝细胞结构被破坏，肝索结构紊乱，出现明显的脂肪空 泡，大量脂滴集聚。与模型组比较，各给药组小鼠肝脏组织 TG、MDA、LDL-C、AST、ALT 水平显著降低 （P＜0.05，P＜0.01，P＜0.001），HDL-C 水平明显升高（P＜0.05）；肝脏组织中肝索结构明显有序，肝细胞空 间缩小，脂肪空泡与脂滴数量减少。（2）与对照组比较，模型组小鼠肠道菌群群落结构存在差异；肠道厚壁菌门 丰度升高，放线菌门丰度降低；肠道放线菌门的红蝽杆菌属（Coriobacteriaceae_UCG-002）丰度显著降低，鼠杆 菌科（Muribaculaceae）的丰度显著升高，差异具有统计学意义（P＜0.05，P＜0.01）。与模型组比较，甲嘎松汤高 剂量组小鼠的菌群结构趋近于对照组，甲嘎松汤可通过重塑菌群相对丰度，逆转酒精诱导的菌群失调；肠道厚 壁菌门丰度明显降低，放线菌门丰度明显升高；肠道的 Coriobacteriaceae_UCG-002 丰度显著升高，Muribaculaceae 的丰度显著降低，差异具有统计学意义（P＜0.05，P＜0.01）。（3）对照组 vs. 模型组与模型组 vs. 甲嘎松汤高剂量 组的差异代谢物取交集，共得到 275 种共有差异代谢物，显著富集于色氨酸代谢通路。相关性分析揭示甲嘎松 汤可通过调控肠道菌群，影响色氨酸通路差异代谢物水平。网络药理学分析筛选出 28 个核心靶点，核心成分 中去氢二异丁香酚的度值最高；联合代谢组学分析得到核心靶点 CYP1A1、MAOA、MAOB。分子对接表明去 氢二异丁香酚与上述 3 个靶点结合稳定。（4）与对照组比较，模型组细胞 TG、TC、MDA、AST、ALT 水平均显 著升高（P＜0.05，P＜0.01），MAOA mRNA 表达水平明显降低（P＜0.05）。与模型组比较，去氢二异丁香酚低、 中、高剂量组细胞 TG、TC、MDA、AST、ALT 水平均显著降低（P＜0.05，P＜0.01），MAOA mRNA 表达水平显著升高（P＜0.01）。结论 甲嘎松汤能改善酒精性肝病小鼠肝脏的脂质累积、氧化应激及肝脏组织病理损伤， 改善其肝功能，可能与通过去氢二异丁香酚等活性成分调节肠道菌群及色氨酸代谢紊乱有关。

Objective To investigate the effects and mechanism of Tibetan medicine Jiagasong Decoction on alcoholic liver disease （ALD） in mice based on gut microbiota，fecal metabolomics，and network pharmacology. Methods （1）An ALD mouse model was induced by intragastric administration of an ethanol liquid diet for 28 days. C57BL/6 mice were randomly divided into control，model，silymarin，and low-，medium-，and high-dose Jiagasong Decoction groups （n=6 per group）. One week after model induction began， drug administration via gavage was initiated. The low- ， medium-，and high-dose Jiagasong Decoction groups received 0.17，0.33，and 0.66 g·kg-1 of Jiagasong Decoction decoction， respectively， while the silymarin group received 50 mg·kg-1 ， once daily for 21 consecutive days. Biochemical methods were used to measure liver tissue levels of triglycerides （TG），malondialdehyde （MDA），lowdensity lipoprotein cholesterol （LDL-C），high-density lipoprotein cholesterol （HDL-C），aspartate aminotransferase （AST），and alanine aminotransferase （ALT）. Lipid accumulation in liver tissue was observed using Oil Red O staining， and histopathological changes were examined via HE staining. 16S rRNA gene sequencing of gut microbiota and metabolic profiling of fecal samples were performed.（2） Network pharmacology was used to predict potential therapeutic targets of Jiagasong Decoction for ALD，and core active components and core targets were screened. Metabolomic and network pharmacology analyses were combined to construct a "differential metabolite-reaction-enzyme-gene" network， and key targets in the tryptophan metabolism pathway were identified and intersected with the core targets from network pharmacology. Molecular docking was performed to validate the interactions between the core active component dehydrodiisoeugenol and the core targets CYP1A1，MAOA，and MAOB.（3） AML-12 cells were treated with low-， medium-，and high-doses of dehydrodiisoeugenol （6.25，12.5，25 µmol·L-1 ） for 24 hours，followed by induction with 400 mmol·L-1 ethanol for 2 hours. Biochemical methods were used to measure cellular TG，total cholesterol （TC）， MDA，ALT，and AST levels. qRT-PCR was employed to assess mRNA expression levels of CYP1A1，MAOA，and MAOB. Results （1） Compared with the control group，the model group showed significantly elevated liver tissue levels of TG，MDA，LDL-C，AST，and ALT （P＜0.05，P＜0.01，P＜0.001） and significantly reduced HDL-C levels （P＜0.001）. Liver tissue displayed disrupted hepatocyte structure， disorganized hepatic cords， prominent fatty vacuoles，and extensive lipid droplet accumulation. Compared with the model group，all treatment groups exhibited significantly reduced liver tissue levels of TG，MDA，LDL-C，AST，and ALT （P＜0.05，P＜0.01，P＜0.001） and significantly increased HDL-C levels （P＜0.05）. Liver tissue showed more orderly hepatic cord structures， reduced hepatocyte space，and decreased numbers of fatty vacuoles and lipid droplets.（2） Compared with the control group， the model group exhibited differences in gut microbiota community structure：an increase in Firmicutes and a decrease in Actinobacteria. The abundance of Coriobacteriaceae_UCG-002（Actinobacteria） was significantly reduced， while the abundance of Muribaculaceae was significantly increased， with statistical significance （P＜0.05， P＜0.01）. Compared with the model group， the high-dose Jiagasong Decoction group displayed a microbiota structure more similar to the control group. Jiagasong Decoction reversed ethanol-induced dysbiosis by reshaping the relative abundance of microbiota： the abundance of Firmicutes was significantly reduced， while that of Actinobacteria was significantly increased. The abundance of Coriobacteriaceae_UCG-002 was significantly elevated， and that of Muribaculaceae was significantly reduced，with statistical significance （P＜0.05，P＜0.01）.（3） Intersection analysis of differential metabolites between the control vs. model groups and the model vs. high-dose Jiagasong Decoction groups identified 275 shared differential metabolites，which were significantly enriched in the tryptophan metabolism pathway. Correlation analysis revealed that Jiagasong Decoction could influence the levels of differential metabolites in the tryptophan pathway by modulating the gut microbiota. Network pharmacology analysis identified 28 core targets，with dehydrodiisoeugenol having the highest degree value among core components. Integrated metabolomic analysis identified the core targets CYP1A1，MAOA，and MAOB. Molecular docking indicated stable binding of dehydrodiisoeugenol to these three targets.（4） Compared with the control group，the model group showed significantly increased cellular levels of TG，TC，MDA，AST，and ALT （P＜0.05，P＜0.01） and significantly decreased MAOA mRNA expression （P＜ 0.05）. Compared with the model group，the low-，medium-，and high-dose dehydrodiisoeugenol groups exhibited significantly reduced cellular levels of TG， TC， MDA， AST， and ALT （P＜0.05， P＜0.01） and significantly increased MAOA mRNA expression （P＜0.01）. Conclusion Jiagasong Decoction can ameliorate lipid accumulation， oxidative stress， and histopathological damage in the liver， and improve liver function in mice with alcoholic liver disease. This effect may be associated with the regulation of gut microbiota and tryptophan metabolic disorders by active components such as dehydrodiisoeugenol.

R285.5

国家自然科学基金项目（8237140578）。