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中国医药导刊 ›› 2025, Vol. 27 ›› Issue (8): 821-831.

• 中医中药 • 上一篇    下一篇

基于网络药理学和实验验证探索荷叶治疗代谢相关脂肪性肝病的机制

任前军1, 李素1, 陈雨晴1, 孟效祖1, 陆炜琴1, 刘旭东2, 赵晓芳2*   

  1. 1.广西中医药大学,广西 南宁 530200;2.广西中医药大学附属瑞康医院,广西 南宁 530011
  • 收稿日期:2025-02-23 修回日期:2025-05-25 接受日期:2025-10-10 出版日期:2025-08-28 发布日期:2025-10-13
  • 基金资助:

    广西自然科学基金项目(2024JJA141239);国家自然科学基金项目(8216140161);广西研究生教育创新计划项目(YCSW2023380)

Mechanism of Lotus Leaves in the Treatment of Metabolic Associated Fatty Liver Disease Based on Network Pharmacology and Experimental Validation

REN Qianjun1, LI Su1, CHEN Yuqing1, MENG Xiaozu1, LU Weiqin1, LIU Xudong2, ZHAO Xiaofang2*   

  1. 1.Guangxi University of Chinese Medicine Guangxi Nanning 530200, China
    2.Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine Guangxi Nanning 530011, China
  • Received:2025-02-23 Revised:2025-05-25 Accepted:2025-10-10 Online:2025-08-28 Published:2025-10-13
  • Supported by:

摘要:

目的:通过网络药理学探索中药荷叶治疗代谢相关脂肪性肝病(metabolic dysfunction associated fatty liver diseaseMAFLD)的潜在作用机制并进行验证。方法:使用中药系统数据库及分析平台(traditional Chinese medicine systems pharmacology database and analysis platformTCMSP)、Uniprot数据库搜集和匹配中药荷叶的活性成分和相应靶点,采用人类基因综合数据库(GeneCards)、人类孟德尔遗传(online Mendelian inheritance in manOMIM)数据库预测MAFLD靶点基因,利用在线交集工具(Venny 2.1)获取药物与疾病的交集靶点;使用String数据库构建蛋白质相互作用关系网络,导出文件后在软件(Cytoscape 3.7.2)中输出核心靶点和构建“中药成分-靶点-疾病”可视化网络图;将核心靶点进行基因本体论(gene ontologyGO)及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomesKEGG)功能和通路的富集分析;使用国家生物技术信息中心(national center of biotechnology informationNCBI)数据库中基因表达矩阵数据进行核心基因的差异分析及回归曲线分析验证,根据受试者工作特征曲线(receiver operating characteristicROC)结果筛选差异显著的基因进行分子对接以评估成分与靶点基因的稳定性;同时,通过成分干预人肝癌细胞株G2型(hepatocellular carcinomas G2HepG2)细胞脂肪变性模型后测定相关蛋白的表达水平。结果:共获得中药荷叶活性成分14个,包括槲皮素、儿茶酸、异鼠李亭等;成分所对应的靶点有177个,疾病对应的靶点有274个,“药物-疾病”交集靶点9个;核心靶点在String数据库中的蛋白相互作用表明荷叶成分可能作用于白细胞介素-6interleukin-6IL6)、白细胞介素-10interleukin-10IL10)、表皮生长因子受体(epidermal growth factor receptorEGFR)、热休克蛋白-β1heat shock protein beta-1HSPB1)、中性粒细胞胞浆因子1neutrophil cytosolic factor 1NCF1)、过氧化物酶体增殖物激活受体αperoxisome proliferator-activated receptor αPPARA)、过氧化物酶体激活物受体δperoxisome proliferator activated receptor δPPARD)、血清对氧磷酶(paraoxonasePON1)、脂肪酸合成酶(fatty acid synthaseFASN)等多个靶点;基因的信号通路分析表明,核心基因可能参与调控FoxO、酪氨酸激酶-信号传导子及转录激活子(Janus tyrosine kinase-signal transducer and activator of transcriptionJAK-STAT)、过氧化物酶体增殖物激活受体(peroxisome proliferators activated receptorsPPAR)、基因组丝氨酸和苏氨酸的蛋白激酶(mitogen-activated protein kinaseMAPK)、肠道炎症等相关信号通路;基因表达数据分析表明在对照组与实验组中最具有显著性的基因是IL6IL10FASNPPARA等,其中只有IL6IL10的曲线下面积(area under the curveAUC)在90%以上;通过分子对接分析得到成分槲皮素与IL6IL10的稳定性较好;细胞实验结果显示不同浓度的槲皮素在干预HepG2细胞脂肪变性模型后,蛋白IL6的表达水平呈浓度依赖性降低。结论:中药荷叶治疗MAFLD的炎症及脂质沉积,其机制可能与槲皮素调控IL6IL10等蛋白表达有关。

  

关键词: 荷叶, 槲皮素, 代谢相关脂肪性肝病(MAFLD), 网络药理学, 白细胞介素-6IL6

Abstract:

Objective: To explore the potential mechanism of action and validation of the Chinese medicine lotus leaves for the treatment of metabolic associated fatty liver disease MAFLD through network pharmacology.Methods: The TCM systems pharmacology database and analysis platform TCMSP and Uniprot database were used to collect and match the active ingredients and corresponding targets of the TCM lotus leaf. MAFLD target genes were predicted using the Human Gene Comprehensive Database GeneCards), and the Human Mendelian Inheritance OMIM database. The online intersection tool Venny 2.1 was used to obtain drug-disease intersection targets. The protein interaction network was constructed using the String databaseand the core targets were exported to the software Cytoscape 3.7.2 for the construction of a visual network diagram of "TCM ingredient-target-disease". The core targets were subjected to gene ontology GO and Kyoto encyclopedia of genes and genomes KEGG functional and pathway enrichment analyses. Differential and regression curve analyses of the core genes were validated using gene expression matrix data from the national center for biotechnology information NCBI databaseand genes with significant differences were screened for molecular docking based on the results of the subjects' operating characteristic curves ROCs to assess the stability of the composition and target genes. Meanwhilethe expression levels of relevant proteins were determined after intervention of human hepatocellular carcinoma cell line G2 type HepG2 cell steatosis model by the components.Results: A total of 14 active ingredients including quercetin catechuic acid isorhamnetin etc. were obtainedwith 177 targets corresponding to the ingredients274 targets corresponding to the diseases and 9 "drug-disease" intersection targets. The protein interactions of the core targets in the String database indicated that the components of Lotus leaves may act on interleukin-6 IL6), interleukin-10 IL10), epidermal growth factor receptor EGFR), heat shock protein-β1 HSPB1), neutrophil cytoplasmic factor 1 NCF1), peroxisome proliferator activated receptor α PPARA), peroxisome proliferator activated receptor αperoxisome proliferators activated receptors PPAR), peroxisome proliferator activated receptor δ PPARD),serum paraoxonase PON1),fatty acid synthase FASN),and several other targets. The signalling pathways of the genes suggest that the core genes may be involved in the regulation of FoxO Janus tyrosine kinase-signal transducer and activator of transcription JAK-STAT),peroxisome proliferator activated receptor PPAR), mitogen-activated protein kinase MAPK), intestinal inflammation and other related signalling pathways. Analysis of gene expression data showed that the most significant genes in the control and experimental groups were IL6 IL10 FASN PPARA etc. of which only IL6 and IL10 had an area under the curve AUC area of more than 90%. The stability of the component quercetin with IL6 and IL10 was obtained by molecular docking analysis. The results of cellular experiments showed that different concentrations of quercetin decreased the expression level of protein IL6 in a concentration-dependent manner after intervening in the HepG2 cell steatosis model.Conclusion: The mechanism of the herbal medicine lotus leaves for the treatment of inflammation and lipid deposition in MAFLD may be related to the regulation of protein expression of IL6 and IL10 through quercetin.


Key words: Lotus leaves , Quercetin , Metabolic associated fatty liver disease , MAFLD); , Network pharmacology , Interleukin-6 , IL6

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