目的 采用网络药理学和动物实验相结合的方法探讨消积丸治疗动脉粥样硬化（atherosclerosis，AS）的作用机制。方法 通过网络药理学进行活性成分-靶点网络构建、蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络分析、基因本体（gene ontology，GO）功能和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。50只雄性ApoE^−∕−小鼠随机分为对照组、模型组、阿托伐他汀（1.3 mg/kg）组和消积丸低、高剂量（8.125、32.500 g/kg）组，模型组和各给药组以高脂饮食喂养，给予相应药物干预4周后，采用全自动生化仪分析小鼠血清脂质水平；采用ELISA法检测小鼠血清中白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和氧化低密度脂蛋白（oxidized low density lipoprotein，ox-LDL）水平；采用苏木素-伊红（HE）、Masson和油红O染色观察小鼠主动脉窦病理变化；采用Western blotting法检测小鼠主动脉组织中TNF-α/κB抑制因子激酶β（inhibitor κB kinase β，IKKβ）/核因子-κB（nuclear factor-κB，NF-κB）信号通路相关蛋白及CD36蛋白表达。结果 共筛选出消积丸136个活性成分及277个作用靶点，4481个AS疾病相关靶点，消积丸和AS的共同靶点有205个，核心靶点主要包括TNF、IL-6、NF-κB、IL-1β等。GO功能和KEGG通路富集分析发现，消积丸治疗AS主要涉及炎症反应、一氧化氮生物合成等过程，调控癌症通路、乙肝通路、TNF信号通路等；在TNF信号通路中进一步筛选得到TNF-α/IKKβ/NF-κB信号通路。动物实验结果显示，与对照组相比，模型组小鼠血清中三酰甘油（triglyceride，TG）、总胆固醇（total cholesterol，TC）、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）、IL-1β、IL-6、TNF-α、ox-LDL水平均显著升高（P＜0.01）；主动脉中TNF-α蛋白表达以及IKKβ、NF-κB p65磷酸化水平均显著升高（P＜0.01）；主动脉窦内膜不光滑，出现大量胶原纤维和脂质沉积（P＜0.01），形成明显的AS斑块。与模型组相比，消积丸组小鼠血清中TC、TG、LDL-C、TNF-α、IL-6、ox-LDL水平均显著降低（P＜0.05、0.01），血清中HDL-C水平显著升高（P＜0.01）；主动脉中TNF-α、CD36蛋白表达以及IKKβ、NF-κB p65磷酸化水平均显著降低（P＜0.05、0.01）；主动脉窦AS斑块减少，胶原蛋白和脂质沉积等情况均得到改善（P＜0.05、0.01）。结论 消积丸能够通过下调TNF-α/IKKβ/NF-κB信号通路相关蛋白表达，减轻炎症反应，调节脂质代谢，从而发挥治疗AS的作用。

Objective To explore the mechanism of Xiaoji Pills (消积丸, XJP) in the treatment of atherosclerosis (AS) by combining network pharmacology and animal experiments.Methods Through network pharmacology, active ingredient-target network construction, protein-protein interaction (PPI) network analysis, gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed. Fifty male ApoE^−∕− mice were randomly divided into control group, model group, atorvastatin (1.3 mg/kg) group and XJP low-, high-dose (8.125, 32.500 g/kg) groups, model group and each administration group were fed with high-fat diet, and after 4 weeks of corresponding drug intervention, lipid levels in serum were analyzed by automatic biochemical analyzer; ELISA method was used to detect the levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and oxidized low density lipoprotein (ox-LDL) in serum. Hematoxylin-eosin (HE), Masson and oil red O staining were used to observe the pathological changes of aortic tissues. Western blotting method was used to detect the expressions of TNF-α/inhibitor κB kinase β (IKKβ)/nuclear factor-κB (NF-κB) signaling pathway related proteins and CD36 protein in aortic tissue. Results A total of 136 active ingredients and 277 targets of XJP, 4481 disease-related targets of AS, and 205 common targets of XJP and AS were screened out, core targets mainly involving TNF, IL-6, NF-κB and IL-1β. GO function and KEGG pathway enrichment analysis showed that the treatment of AS by XJP mainly involved inflammatory reaction, nitric oxide biosynthesis and other processes, regulating cancer pathway, hepatitis B pathway and TNF signaling pathway. TNF-α/IKKβ/NF-κB signaling pathway was further screened in TNF signaling pathway. The results of animal experiments showed that compared with control group, levels of triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), IL-1β, IL-6, TNF-α, ox-LDL in serum of mice in model group were significantly increased (P < 0.01), TNF-α protein expression, IKKβ and NF-κB p65 phosphorylation levels in aorta were significantly increased (P < 0.01), intima of aortic sinus was not smooth with a large number of collagen fibers and lipid deposition (P < 0.01), and obvious AS plaques were formed. Compared with model group, levels of TC, TG, LDL-C, TNF-α, IL-6 and ox-LDL in serum of mice in XJP groups were significantly decreased (P < 0.05, 0.01). TNF-α, CD36 protein expressions and IKKβ, NF-κB p65 phosphorylation levels in aorta were significantly decreased (P < 0.05, 0.01); aortic sinus AS plaques were decreased, collagen protein and lipid deposition were improved (P < 0.05, 0.01). Conclusion XJP can treat AS by downregulating related proteins expression in TNF-α/IKKβ/NF-κB signaling pathway, reducing inflammation and regulating lipid metabolism.

R285.5

国家自然科学基金资助项目（81673848）