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目的：基于网络药理学并通过实验验证研究温脾消导方对非酒精性脂肪肝病（non-alcoholic fatty liver disease,NAFLD）大鼠的治疗作用，以及对SIRT1-FoxO1-自噬通路的改变及分子机制。方法：通过TCMSP、BATMAN-TCM、GeneCards和OMIM数据库获取温脾消导方治疗NAFLD的潜在活性成分及作用靶点，使用Cytoscape 3.9.1软件和STRING数据库构建中药-活性成分-疾病靶点网络、蛋白质-蛋白质相互作用（PPI）网络并筛选关键活性成分和核心靶点，通过R4.2.2软件对交集靶点进行GO和KEGG富集分析。通过高脂饲料喂养大鼠12周建立NAFLD模型，造模成功后用温脾消导方低（7.43 g/kg）、中（14.85g/kg）、高（29.70 g/kg）剂量和多烯磷脂酰胆碱胶囊干预8周。采用HE染色、油红O染色、Western Blot等方法进行动物体内药效验证和机制探讨。结果：网络药理学预测结果表明，温脾消导方可能通过槲皮素、甲基庚烯酮、木犀草素和山奈酚等关键成分和STAT3、JUN、MYC、ESR1等核心靶点治疗NAFLD;KEGG富集分析显示温脾消导方可能通过调控AGE-RAGE、IL-17、FoxO和TNF信号通路等治疗NAFLD。动物实验验证结果表明，温脾消导方低、中、高剂量和多烯磷脂酰胆碱能够减轻NAFLD大鼠肝脏脂肪变性和脂滴积累程度，显著降低血清中TC、TG、LDL-C、AST和ALT的含量（P<0.05），升高HDL-C的含量（P<0.05），显著降低肝脏中Ac-FoxO1、p62蛋白表达水平（P<0.05），显著升高FoxO1、LC3Ⅱ/LC3Ⅰ、SIRT1蛋白表达水平（P<0.05）。结论：温脾消导方能够显著改善NAFLD大鼠肝功能、血脂水平和肝脏脂肪变性，其机制可能与激活SIRT1-FoxO1-自噬通路有关。
Abstract：Objective: To study the therapeutic effect of Wenpi Xiaodao Decoction on non-alcoholic fatty liver disease(NAFLD) in rats based on network pharmacology and experimental verification, and the changes of SIRT1-FoxO1-autophagy pathway and its molecular mechanism. Methods: TCMSP, BATMAN-TCM, GeneCards and OMIM databases were used to obtain the potential active ingredients and targets of Wenpi Xiaodao Decoction in treating NAFLD. Cytoscape 3.9.1 software and STRING database were used to construct TCM-active ingredient-disease target network and protein protein interaction(PPI) network, and key active ingredients and core targets were screened. GO and KEGG enrichment analysis of intersection targets was performed by R4.2.2 software. NAFLD model was established by feeding SD rats with high-fat diet for 12 weeks. After successful modeling, the rats were intervened with low(7.43 g/kg), medium(14.85 g/kg), high(29.70 g/kg) doses of Wenpi Xiaodao Decoction and polyene phosphatidylcholine capsules for 8 weeks. HE staining, oil red O staining, Western blot and other methods were used to verify the efficacy in vivo and explore the mechanism. Results: The prediction results of network pharmacology showed that Wenpi Xiaodao Decoction may treat NAFLD through key components such as quercetin, Methylheptenone, luteolin, kaempferol and core targets such as STAT3, JUN, MYC, ESR1. KEGG enrichment analysis showed that Wenpi Xiaodao Decoction may treat NAFLD by regulating AGE-RAGE, IL-17, FoxO and TNF signaling pathways. The results of animal experiments showed that low, medium and high doses of Wenpi Xiaodao Decoction and polyene phosphatidylcholine could reduce the degree of liver steatosis and lipid droplet accumulation in NAFLD rats, significantly reduce the contents of TC, TG, LDL-C, AST and ALT in serum(P<0.05), increase the content of HDL-C(P<0.05), significantly reduce the expression levels of ac-foxo1and p62 protein in liver(P<0.05), and significantly increase the levels of FoxO1, LC3 Ⅱ/LC3 Ⅰ SIRT1 protein expression level(P<0.05). Conclusion: Wenpi Xiaodao Decoction can significantly improve liver function, blood lipid levels and liver steatosis.Its mechanism may be related to the activation of SIRT1-FoxO1-autophagy pathway.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20240429.002
中图分类号:R285.5
引用信息:
[1]高松林,韦柳婷,覃雁,等.基于网络药理学及实验验证探讨温脾消导方治疗非酒精性脂肪肝病的作用机制[J].海南医学院学报,2024,30(18):1403-1413.DOI:10.13210/j.cnki.jhmu.20240429.002.
基金信息:
广西研究生教育创新计划项目（YCBZ2023148）; 广西科技基地和人才专项（桂科AD19245168）; 国医大师黄瑾明学术思想与临床诊疗传承发展研究中心建设项目[桂中医大党（2022）24号]; 黄贵华桂派中医大师培养项目[桂中医药科教发（2022）6号]; 黄贵华广西名中医传承工作室项目[桂卫中医发（2017）2号]~~
2024-04-29
2024-04-29
2024-04-29