基于网络药理学分析灯盏乙素治疗动脉粥样硬化的分子机制和体内验证

Network Pharmacology-based Analysis of the Antiatherosclerosis Mechanism of Scutellarin and Experimental Validation in Vivo

目的基于网络药理学分析灯盏乙素(Scutellarin)治疗动脉粥样硬化(atherosclerosis,AS)潜在的分子生物学机制。方法通过PubChem数据库进行灯盏乙素潜在的靶点预测,运用Disgenet,CeneCards筛选与AS关联的靶基因,利用Cytoscape 3.9.0建立疾病-靶点-成份可视化网络,使用String构建蛋白互作网络图,利用David数据库进行GO及KECG的网络构建。另外,高脂喂养复制APOE^(-/-)小鼠AS模型,用不同浓度的灯盏乙素灌胃治疗,利用RT-PCR方法验证筛选所得核心通路的表达变化。结果(1)网络药理学预测结果:灯盏乙素治疗AS的核心靶点共有肿瘤坏死因子-α(TNF-α),AKT丝氨酸/苏氨酸激酶(AKT serine/threonine kinase,MAPK),低密度脂蛋白受体(Low density lipoprotein receptor),载脂蛋白E(apolipoprotein E,APOE)等共106个。灯盏乙素调控PI3K/AKT信号通路(hsa04151)、Ras信号通路(hsa04014)、MAPK信号通路(hsa04010)(P<0.05)等涉及细胞的增殖,炎症反应,氧化应激,及细胞凋亡等生物学过程来治疗AS;(2)体内实验验证的结果:和模型组相比,灯盏乙素组呈浓度依耐性地下调PI3K、AKT、mTOR和Bcl-2的mRNA表达,上调Bax和Caspase-3的mRNA表达,(P<0.05)。另外,和模型组相比,灯盏乙素则呈浓度依耐性地下调小鼠血清TNF-α和IL-1β的含量(P<0.05)。结论经过网络药理学筛选和RT-PCR验证,灯盏乙素通过调控PI3K/AKT/mTOR信号通路抑制APOE^(-/-)小鼠AS模型炎症反应,促进细胞凋亡而治疗AS。

Objective To analyze the potential molecular biological mechanism of scutellarin for the treatment of atherosclerosis(AS)based on network pharmacology.Methods The PubChem database was used to predict the potential targets of scutellarin,and the Disgenet and CeneCards databases were used to predict and screen the genes related to atherosclerosis,and the Cytoscape 3.7.0 software was used to establish and construct the component-target-disease network visualization network.The GO and KECG pathways were analyzed using the David database.In addition,an APOE^(-/-)mouse AS model was developed and treated with different concentrations of scutellarin,and the expression changes of the core targets were verified by RT-PCR.Results 1.Network pharmacological prediction results:the core targets of Scutellarin for AS treatment were tumor necrosis factor-α(TNF-α),AKT serine/threonine kinase 1(MAPK),low density lipoprotein receptor(LDLR),apolipoprotein E(APOE),among other targets,with 106 in total.The key targets of Scutellarin are PI3K/AKT signaling pathway(hsa04151),Ras signaling pathway(hsa04014)and MAPK signaling pathway(hsa04010)(P<0.05),which are involved in cell proliferation,inflammatory response,oxidative stress and apoptosis,to treat AS.2.Results of in vivo experiments:compared with the model group,the mRNA expression of PI3K,AKT,mTOR and Bcl-2 was down-regulated and the mRNA expression of Bax and Caspase-3 was up-regulated in the Scutellarin group.In addition,compared with model group,Scutellarin down-regulated the contents of TNF-αand IL-1βin serum concentration-dependent(P<0.05).Conclusion Through network pharmacology screening and RT-PCR verification,Scutellarin inhibits the process of inflammatory response in a mouse model of AS by regulating the PI3K/AKT/mTOR signaling pathway.