目的 通过Meta分析、网络药理学分析及体内实验验证，阐述二陈汤（Erchen Decoction，ECD）发挥燥湿化痰功效治疗非酒精性脂肪性肝病（non-alcoholic fatty liver disease，NAFLD）的临床证据和潜在机制网络与物质基础和靶点验证的现代表征关系。方法 筛选符合纳入标准的ECD治疗NAFLD的临床随机对照研究，进行研究质量评价和数据提取，并运用Revman 5.4软件进行数据分析。筛选ECD和燥湿化痰类中药的活性成分及对应靶点，运用Cytoscape 3.10.1等软件构建ECD燥湿化痰功效的“药物-活性成分-关键靶点”作用网络，并将ECD燥湿化痰功效的关键靶点与NAFLD疾病靶点取交集，通过Matescape平台对交集靶点进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。根据网络药理学和现代物质基础研究，构建NAFLD动物模型，检测动物血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨酸转移酶（aspartate aminotransferase，AST）、总胆固醇（total cholesterol，TC）、三酰甘油（triglyceride，TG）水平；苏木素-伊红（hematoxylin-eosin，HE）染色和油红O染色检测肝脏组织病理变化；利用实时定量逆转录聚合酶链反应（real time fluorescence quantitative reverse transcriptase-polymerase chain reaction，qRT-PCR）技术对ECD治疗NAFLD的核心靶点进行实验验证。结果 Meta分析显示，ECD单独或联合其他方法治疗NAFLD的有效率明显高于对照组，且在改善ALT、AST、TC、TG等指标方面均优于对照组。网络药理学研究共获得ECD活性成分145种，相关靶点392个，与燥湿化痰类中药靶点取交集后，得到ECD燥湿化痰功效的关键作用靶点37个，治疗NAFLD涉及过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor gamma，PPARG）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、前列腺素内过氧化物合酶2（prostaglandin G/H synthase 2，PTGS2）、肿瘤蛋白p53（tumor protein p53，TP53）、糖原合酶激酶-3β（glycogen synthase kinase-3 beta，GSK-3β）等25个关键靶点。富集分析揭示了545个生物学过程、13个细胞组成组分、40个分子功能和136条KEGG通路。其信号通路主要涉及脂质和动脉粥样硬化、非酒精性脂肪肝、细胞凋亡、磷脂酰肌醇-3-羟激酶（phosphatidylinositol-3-hydroxykinase，PI3K）-蛋白激酶B（protein kinase B，Akt）信号通路、丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）信号通路、肿瘤坏死因子（tumor necrosis factor，TNF）信号通路白细胞介素-17（interleukin-17，IL-17）信号通路等。动物实验结果表明，ECD可以显著降低NAFLD小鼠血脂水平，改善肝功能异常，上调小鼠肝脏中PPARG和Bcl-2 mRNA的表达。结论 ECD对NAFLD具有一定的治疗作用，表现为发挥燥湿化痰功效，降低ALT、AST、TC、TG水平，改善肝细胞脂肪变性和积聚，改善右胁不适或胀闷、周身困重、大便黏滞不爽等证候，其功效作用网络与PPARG、Bcl-2、PTGS2等多个关键靶点以及脂质和动脉粥样硬化、非酒精性脂肪肝、细胞凋亡等信号通路有关，共同发挥调节脂质代谢、改善肝细胞凋亡的作用。结合动物实验发现PPARG和Bcl-2可能是ECD发挥燥湿化痰功效治疗NAFLD的核心靶点。

Objective To elucidate the clinical evidence and potential mechanism of Erchen Decoction (二陈汤, ECD) in the treatment of non-alcoholic fatty liver disease (NAFLD) through Meta-analysis combined with network pharmacology analysis and in vivo experiments verification. Methods Randomized controlled clinical studies of ECD in the treatment of NAFLD that met the inclusion criteria were screened, study quality evaluation and data extraction were conducted, and data analysis was performed using Revman 5.4 software. The active components and their corresponding targets from ECD and traditional Chinese medicines (TCMs) with dampness and phlegm elimination were screened. The “drug-active ingredient-key target” action network for the dampness and phlegm elimination efficacy of ECD was constructed by using Cytoscape 3.10.1 and other software, and the key target of the dampness and phlegm elimination efficacy of ECD was intersected with the disease target of NAFLD. The gene ontology (GO) function and the Kyoto encyclopedia of genes and genomes (KEGG) enrichment of the intersection targets were analyzed using the Matescape platform. According to the network pharmacology and modern pharmacodynamic substance research, the animal model of NAFLD was constructed. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC) andtriglyceride (TG) in serum were detected. The pathological changes of liver tissue were detected by hematoxylin-eosin (HE) staining and oil red O staining. And the core target of ECD in the treatment of NAFLD was verified by real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) technology. Results Meta-analysis showed that ECD had superior improvements in key indicators such as ALT, AST, TC and TG compared to the control group. A total of 145 active ingredients and 392 related targets were obtained from network pharmacology, and 37 key targets were obtained for the dampness and phlegm elimination effect of ECD, and 25 key targets such as peroxisome proliferator-activated receptor gamma (PPARG), B-cell lymphoma-2 (Bcl-2), prostaglandin G/H synthase 2 (PTGS2), tumor protein p53 (TP53), glycogen synthase kinase-3 beta (GSK-3β) were involved in the treatment of NAFLD. Enrichment analysis revealed 545 biological processes, 13 cell components, 40 molecular functions and 136 KEGG pathways. The signaling pathways are mainly related to lipid and atherosclerosis, non-alcoholic fatty liver disease, apoptosis, phosphatidylinositol-3-hydroxykinase-protein kinase B (PI3K-Akt) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, tumor necrosis factor (TNF) signaling pathway, interleukin-17 (IL-17) signaling pathway, etc. The results of animal experiments showed that ECD could effectively reduce the levels of blood lipids and improve the liver function in NAFLD mice, and up-regulate the expression of PPARG and Bcl-2 mRNA in the livers of mice. Conclusion ECD has a certain therapeutic effect on NAFLD, manifested by its ability to exert dampness and phlegm elimination effects, reducing levels of ALT, AST, TC, and TG, ameliorating the accumulation and degeneration of hepatocyte fat. It also improves symptoms such as right hypochondrium discomfort or swelling, heavy body, and unpleasant stool. The efficacy is associated with a network of key targets including PPARG, Bcl-2, PTGS2 and multiple signaling pathways such as lipid and atherosclerosis,non-alcoholic fatty liver disease and apoptosis. These collectively play a role in regulating lipid metabolism and reducing hepatocyte apoptosis. Combined with animal experiments, it was found that PPARG and Bcl-2 may be the core targets through which ECD exerts its dampness and phlegm elimination effects in the treatment of NAFLD.

R285.64;R285

国家重点研发计划项目（2022YFC3502100）;山东大学高等医学研究院前沿学科发展基金（GYY2023QY01）