摘要
目的 基于网络药理学、转录组学及分子对接探讨四君子汤抗动脉粥样硬化(Atherosclerosis,AS)的作用靶点及分子机制。方法 借助TCMSP等数据库挖掘四君子汤活性成分及动脉粥样硬化靶点。构建“药物-成分-靶点-疾病”网络和蛋白质–蛋白质相互作用(Proteinprotein interaction,PPI)网络,进行基因本体(Gene ontology,GO)功能及京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。对小鼠主动脉进行转录组学测序,筛选差异表达基因(Differential gene expression,DEG);与网络药理学进行联合分析并进行分子对接。采用全自动生物化学分析仪检测甘油三酯(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)含量;苏木精-伊红(Hematoxylin and eosin,HE)染色和油红O染色观察主动脉病理变化情况;采用实时定量逆转录聚合酶链反应(Quantitative reverse transcription polymerase chain reaction,qRT-PCR)、蛋白质印迹法(Western blotting)法验证表皮生长因子(Epidermal growth factor,EGF)、OLR1、分泌型磷蛋白1(Secretory phosphoprotein 1,SPP1)的基因及蛋白表达水平;采用酶联免疫吸附测定(Enzyme-linked immunosorbent assaynzyme-linked immunosorbent assay,ELISA)检测血清丙二醛(Malondiadehyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)含量。结果 网络药理学筛选出四君子汤核心活性成分(槲皮素、山柰酚等)和核心靶点(AKT1、IL-6等),作用机制可能与PI3K-Akt、MAPK等信号通路相关。转录组学分析发现动脉粥样硬化模型组存在339个DEGs,四君子汤干预能有效逆转其基因的表达。联合分析确定核心靶点为EGF、OLR1和SPP1。分子对接表明,山柰酚、槲皮素等与上述靶点具有良好亲和力。动物实验证实,与正常组相比,动脉粥样硬化模型组小鼠血清TC、TG和LDL-C水平升高(P<0.01),HDL-C水平降低(P<0.01),主动脉斑块面积和脂质沉积显著增加,氧化应激MDA水平升高,SOD水平显著降低(P<0.01),且EGF、OLR1、SPP1的mRNA和蛋白表达均显著上调(P<0.01)。经四君子汤治疗后,上述指标均得到显著改善。结论 四君子汤可能通过山柰酚、槲皮素和柚皮素等活性成分作用于OLR1、SPP1和EGF等核心靶点,调节脂质代谢,抑制氧化应激,从而发挥抗动脉粥样硬化作用。
Objective To explore the therapeutic targets and molecular mechanisms of Sijunzi Decoction(SJZD)against atherosclerosis(AS)based on network pharmacology,transcriptomics,and molecular docking.Methods Active components of Sijunzi Decoction and atherosclerosis(AS)targets were identified using databases such as TCMSP.A“drug-component-target-disease”network and a protein-protein interaction(PPI)network were constructed,followed by Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.Transcriptomic sequencing of mouse aortic tissues was performed to screen differentially expressed gene(DEG),which were subsequently integrated with network pharmacology findings for conjoint analysis and molecular docking.An automated biochemical analyzer was employed to measure serum levels of triglycerides(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C).Pathological changes in the aorta were observed via hematoxylin-eosin(HE)and oil red O staining.Quantitative realtime PCR(qRT-PCR)and Western blotting were used to validate the mRNA and protein expression levels of EGF,OLR1,and SPP1.Serum malondialdehyde(MDA)and superoxide dismutase(SOD)levels were detected by enzymelinked immunosorbent assay(ELISA).Results Network pharmacology identified core active components of Sijunzi Decoction(e.g.,quercetin,kaempferol)and key targets(e.g.,AKT1,IL-6),suggesting that the mechanism of action may involve signaling pathways such as PI3K-Akt and MAPK.Transcriptomic analysis revealed 339 DEGs in the AS model group,and intervention with Sijunzi Decoction effectively reversed the expression of these genes.Integrated analysis pinpointed EGF,OLR1,and SPP1 as core targets.Molecular docking demonstrated strong binding affinity between kaempferol,quercetin,and the aforementioned targets.Animal experiments confirmed that,compared with the control group,the AS model group exhibited significantly elevated serum TC,TG,and LDL-C levels(P<0.01),reduced HDL-C levels(P<0.01),increased aortic plaque area and lipid deposition,elevated oxidative stress marker MDA,and significantly decreased SOD activity(P<0.01).Moreover,mRNA and protein expression of EGF,OLR1,and SPP1 were markedly up-regulated(P<0.01).Treatment with Sijunzi Decoction significantly ameliorated all these indicators.Conclusion Sijunzi Decoction may exert anti-AS effects by regulating lipid metabolism and suppressing oxidative stress through the actions of active components such as kaempferol,quercetin,and naringenin on core targets including OLR1,SPP1,and EGF.
作者
隋玥含
何会
罗涛红
战丽彬
王莹
SUI Yuehan;HE Hui;LUO Taohong;ZHAN Libin;WANG Ying(Basic Medical College,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Liaoning Key Laboratory of Modern Research on Spleen Visceral Manifestations Theory,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Liaoning Key Laboratory for TCM Spleen-Viscera-State Modern Research,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《世界科学技术-中医药现代化》
北大核心
2026年第2期348-363,共16页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会青年科学基金项目(82305061):基于“脾统血濡脉”探讨健脾益气法调控LOX-1/SPP1/EG通路驱动mtROS预防AS分子机制研究,负责人:王莹
辽宁省科技计划联合计划(基金)项目(2023-MSLH-198):LOX-1/SPP1/EGF通路介导线粒体能量代谢驱动线粒体ROS探讨四君子汤防治动脉粥样硬化的作用及机制研究,负责人:王莹
辽宁省中医药创新团队项目(LNZYYCXTD-CCCX-003):中医“五脏系统”调控生命活动科学原理及创新研究团队,负责人:战丽彬。
关键词
网络药理学
转录组学
分子对接
四君子汤
动脉粥样硬化
Network pharmacology
Transcriptomic techniques
Molecular docking
Sijunzi Decoction
Atherosclerosis
