目的 采用网络药理学、分子对接及实验验证的方法，探讨大黄治疗酒精性肝炎的作用机制，为临床应用提供依据。方法 运用中药系统药理学数据库与分析平台（TCMSP）及查阅国内外相关文献收集大黄活性成分，通过GeneCards、OMIM、TTD数据库收集酒精性肝炎靶点，将成分-疾病靶点取交集基因，运用Cytoscape软件构建大黄-活性成分-酒精性肝炎-靶基因网络图，在STRING平台制作蛋白质-蛋白质相互作用（PPI）网络，通过Metascape平台对交集靶点进行富集分析，获得靶点功能及通路。运用Vina等软件对大黄的关键活性成分与关键靶点进行分子对接。建立酒精性肝炎小鼠模型，设置对照组、模型组、氢化可的松（20.8 mg·kg^-1）组和芦荟大黄素低、中、高剂量（20、40、80 mg·kg^-1）组，观察对各组小鼠肝脏指数影响；试剂盒法测定血清中天冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）水平；苏木精-伊红（HE）染色和油红O染色观察小鼠肝组织病理学以及脂质积累情况；Western blotting检测小鼠肝组织中NOD样受体热蛋白结构域相关蛋白3（NLRP3）、白细胞介素-18（IL-18）、IL-1β、剪切型半胱天冬酶-3（cleaved Caspase-3）、半胱天冬酶-3（Caspase-3）和半胱天冬酶-1（Caspase-1）的蛋白表达。结果 大黄16个活性成分对应50个靶点，酒精性肝炎对应6 888个靶点，二者取交集获得45个靶点。其中IL-1β、CASP3、GSDMD、骨髓细胞瘤癌基因（MYC）、前列腺素内过氧化物合酶2（PTGS2）以及蛋白激酶Cα（PRKCA）与核心成分芦荟大黄素分子对接结果良好。基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析发现，细胞凋亡途径可能是大黄治疗酒精性肝炎的重要途径。体内实验结果显示，芦荟大黄素能够缓解小鼠酒精性肝炎，并显著降低肝脏指数（P＜0.001）和AST、ALT水平（P＜0.05）。HE染色与油红O显示，与模型组相比，芦荟大黄素低、中、高剂量组恢复了肝组织结构损伤，降低了炎症细胞浸润，恢复了肝索排列紊乱状态以及减少了脂滴堆积和脂肪变性。Western blotting结果显示，与模型组相比，氢化可的松组以及芦荟大黄素低、中、高剂量组NLRP3、IL-18、IL-1β、GSDMD、cleaved Caspase-3、Caspase-3和Caspase-1蛋白表达水平均有降低，其中芦荟大黄素中剂量组IL-1β、GSDMD、Caspase-3和Caspase-1蛋白表达水平显著降低（P＜0.05、0.01）。结论 大黄治疗酒精性肝炎可通过多成分、多靶点、多通路途径来发挥作用，其中芦荟大黄素可减轻小鼠酒精性肝炎，其机制可能与抑制Caspase-1、Caspase-3介导的经典细胞焦亡和凋亡通路有关。

Objective To explore the mechanism of Rhei Radix et Rhizoma in the treatment of alcoholic hepatitis by means of network pharmacology, molecular docking and experimental verification, and to provide evidence for the clinical application of Rhei Radix et Rhizoma in the treatment of alcoholic hepatitis.Methods The active components of Rhei Radix et Rhizoma were collected using TCMSP and relevant literature at home and abroad. The targets of alcoholic hepatitis were collected by GeneCards, OMIM and TTD databases, and the intersection genes of constituents and disease targets were selected. The Rhei Radix et Rhizoma-active ingredientalcoholic hepatitis-target gene network map was constructed by Cytoscape software. The protein interaction network was made on the STRING platform. The intersection targets were enriched and analyzed by Metascape platform to obtain the target functions and pathways. The key active components of Rhei Radix et Rhizoma were docked with key targets by Vina and other software. A mouse model of alcoholic hepatitis was established, and control group, model group, cortisone (20.8 mg·kg^-1) group and aloe emodin lowdose, medium-dose and high-dose (20, 40, 80 mg·kg^-1) groups were set up to observe the effects on liver index of mice in each group. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined by kit method. Liver histopathology and lipid accumulation were observed by HE staining and oil red O staining. Western blotting analysis of NLRP3, IL-18, IL-1β, cleaved Caspase-3, Caspase-3, and Caspase-1 protein expression in mouse liver tissues.Results A total of 16 active components of Rhei Radix et Rhizoma correspond to 50 targets, alcohol hepatitis correspond to 6 888 targets, and 45 targets were obtained by intersection of the two. IL-1β, CASP3, MYC, PTGS2 and PRKCA had good docking results with aloe emodin. GO and KEGG results showed that apoptosis pathway may be an important pathway for Rhei Radix et Rhizoma treatment of alcoholic hepatitis. The results of in vivo experiment showed that aloe emodin could alleviate alcoholic hepatitis in mice, and significantly reduce liver index (P<0.001) and AST and ALT levels (P<0.05). HE staining and oil red O showed that, compared with the model group, aloe emodin groups restored the damage of dry tissue structure, reduced inflammatory cell infiltration, restored the disorder of hepatic cord arrangement, and reduced lipid droplet accumulation and steatosis. Western blotting results showed that compared with the model group, protein expression levels of NLRP3, IL-18, IL-1β, GSDMD, cleaved Caspase-3, Caspase-3, and Caspase-1 were decreased in cortisone group and aloe albumin dose groups. The expression levels of IL-1β, GSDMD, Caspase-3 and Caspase-1 protein in aloe emodin medium dose group were significantly decreased (P<0.05).Conclusion Rhei Radix et Rhizoma can treat alcoholic hepatitis through multicomponent, multi-target and multi-pathway pathways, among which aloe emodin can alleviate alcoholic hepatitis in mice, and its mechanism may be related to the inhibition of classic cell pyrodeath and apoptosis pathways mediated by Caspase-1 and Caspase-3.

R285.5

国家自然科学基金联合基金重点支持项目（U21A200211）；国家资助博士后研究人员计划资助（GZC20231759）