目的 采用"系统评价定疗效指标-网络药理学预测靶点-交互燥湿化痰类中药定功效-体内验证疗效指标和功效靶点"的集成创新策略,解析二陈汤发挥燥湿化痰功效治疗高脂血症临床证据和作用靶点的现代表征关系。方法 筛选符合纳入标准的二陈汤治疗高脂血症临床随机对照研究,抽提与二陈汤燥湿化痰功效密切相关的临床证候,继而开展临床研究Meta分析。筛选并收集二陈汤和燥湿化痰类中药活性成分及其对应的靶点,运用Cytoscape 3.9.1等软件构建二陈汤燥湿化痰功效的"药物-活性成分-关键靶点"网络,将二陈汤燥湿化痰功效的关键靶点与高脂血症疾病靶点交集,通过Matescape平台对共同靶点进行基因本体(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。基于网络药理学和现代药效物质研究,通过高脂饮食构建高脂血症模型,利用qRT-PCR技术对二陈汤燥湿化痰功效治疗高脂血症的作用靶点进行验证。结果 Meta分析显示,二陈汤单独或联合其他治疗在治疗高脂血症患者的总胆固醇(total cholesterol,TC)、三酰甘油(triglyceride,TG)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)的疗效方面均优于对照组。网络药理学研究得到二陈汤燥湿化痰功效的关键作用靶点37个,治疗高脂血症涉及过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor gamma,PPARG)、前列腺素内过氧化物合酶2(prostaglandin G/H synthase 2,PTGS2)、雌激素受体1(estrogen receptor 1,ESR1)、肿瘤蛋白p53(tumor protein p53,TP53)、AKT丝氨酸/苏氨酸激酶1(RAC-alpha serine/threonine-protein kinase,AKT1)、PPARA等20个关键靶点。GO分析涉及小分子代谢过程的调节、脂质代谢过程的调节等生物过程,KEGG分析涉及脂质和动脉粥样硬化、脂肪细胞中脂肪分解的调节、血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)信号通路和单磷酸腺苷活化蛋白激酶(AMP-activated protein kinase,AMPK)信号通路等91条通路。体内实验结果表明,二陈汤能够有效改善高脂血症大鼠血脂水平(P<0.05、0.01、0.001),上调高脂血症大鼠肝脏中PPARα和PPARγ mRNA的表达(P<0.05、0.001),下调脂肪组织中PPARγ mRNA的表达(P<0.05、0.01)。结论 二陈汤治疗高脂血症时燥湿化痰功效表征为TC、TG、LDL-C水平下降和HDL-C水平升高,功效作用网络与PPARG、AKT1、PPARA等多个关键靶点和VEGF信号通路、AMPK信号通路有关,共同发挥对脂质代谢、脂肪细胞中脂肪分解调节、肝脂调控等的作用。结合其功效物质基础分析和体内实验发现PPARα和PPARγ是二陈汤发挥燥湿化痰功效治疗高脂血症的核心靶点。

Objective To elucidate the modern evidence-based relationships between the clinical efficacy of Erchen Decoction (二陈汤) in treating hyperlipidemia by eliminating dampness and phlegm and effect target based on integrated innovation strategy of "systematic evaluation to determine the efficacy index-network pharmacological prediction target-interaction of dampness and phlegm-eliminating traditional Chinese medicine to clarify the efficacy index-in vivo experiment to verify the efficacy index and efficacy target". Methods Clinical randomized controlled trials of Erchen Decoction for the treatment of hyperlipidemia that meet the inclusion criteria was selected, clinical syndromes closely related to the efficacy of Erchen Decoction in drying dampness and resolving phlegm was extracted, and then a meta-analysis of clinical studies was conducted. The active ingredients and corresponding targets of Erchen Decoction and traditional Chinese medicine for drying dampness and resolving phlegm were screened and collected, "drug-active ingredient-key target" network for the drying dampness and resolving phlegm effect of Erchen Decoction was construct by Cytoscape 3.9.1 software. The key targets of Erchen Decoction's drying dampness and resolving phlegm effect were intersected with those of hyperlipidemia disease, and gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis on common targets were performed by Matescape platform. Based on network pharmacology and modern pharmacological substance research, a hyperlipidemia model was constructed through a high-fat diet, and qRT-PCR technology was used to verify the therapeutic targets of Erchen Decoction's drying dampness and resolving phlegm effect in the treatment of hyperlipidemia. Results Meta-analysis revealed that Erchen Decoction either as a standalone treatment or in conjunction with other therapies, exhibited superior outcomes compared to the control group in managing total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C). A total of 37 key targets of Erchen Decoction for eliminating dampness and phlegm were obtained from the network pharmacological research. The treatment of hyperlipidemia involved 20 key targets such as peroxisome proliferator-activated receptor gamma (PPARG), prostaglandin G/H synthase 2 (PTGS2), estrogen receptor 1 (ESR1), tumor protein p53 (TP53), RAC-alpha serine/threonine-protein kinase (AKT1) and PPARA. GO analysis involved biological processes such as the regulation of small molecule metabolism and lipid metabolism, while KEGG analysis involved 91 pathways such as lipid and atherosclerosis, regulation of lipolysis in adipocytes, vascular endothelial growth factor (VEGF) signaling pathway and AMP-activated protein kinase (AMPK) signaling pathway. The in vivo experiment results showed that Erchen Decoction could effectively improve the level of blood lipids in hyperlipidemia rats (P<0.05, 0.01, 0.001), up-regulate the expressions of PPARα and PPARγ mRNA in liver (P<0.05, 0.001), and down-regulate the expression of PPARγ mRNA in adipose tissue of hyperlipidemia rats (P<0.05, 0.01). Conclusion The drying dampness and resolving phlegm effectiveness of Erchen Decoction in hyperlipidemia treatment is marked by a reduction in TC, TG and LDL-C levels, coupled with an elevation in HDL-C levels. This efficacy network is closely linked to several key targets including PPARG, AKT1, PPARA, as well as pathways such as VEGF signaling pathway and AMPK signaling pathway. These elements collectively play a role in lipid metabolism, regulation of lipolysis in adipocytes and liver lipid regulation. Combined with a fundamental analysis of its functional components and in vivo experiments, PPARα and PPARγ are the central targets of Erchen Decoction for treating hyperlipidemia, utilizing its dampness elimination and phlegm resolution properties.

国家重点研发计划项目（2022YFC3502100）；济南市科研带头人工作室项目（202228099）；高等医学研究院前沿学科发展基金（GYY2023QY01）