基于网络药理学和分子对接分析二氢血根碱抗氧化的作用机制

Analysis of Antioxidant Mechanism of Dihydrosanguinarine Based on Network Pharmacology and Molecular Docking

目的:检测多刺绿绒蒿中二氢血根碱的含量并探索其抗氧化活性,利用网络药理学和分子对接技术探索二氢血根碱通过抗氧化作用对治疗阿尔茨海默病(AD)、甲状腺相关性眼病(TAO)、放射性肺损伤(RILI)、帕金森病(PD)和支气管肺发育不良(BPD)的作用机制。方法:采用高效液相色谱法对多刺绿绒蒿中二氢血根碱的含量进行测定;通过薄层层析-生物自显影法判定其抗氧化活性;综合利用SuperPred、Genecards和OMIM等在线数据库搜集二氢血根碱的靶点和治疗AD、TAO、RILI、PD、BPD的靶点,构建二氢血根碱治疗疾病的“化合物-靶点-疾病”和蛋白互作网络;基于Metascape数据库及CytoNCA插件进行KEGG通路富集分析;采用Discovery Studio 2019 Client软件对活性成分与潜在靶点进行分子对接验证结合活性。结果:研究显示多刺绿绒蒿中二氢血根碱的含量为0.50 mg/g;薄层层析-生物自显影结果显示二氢血根碱具有抗氧化活性;通过网络药理学分析获得二氢血根碱治疗以上五种疾病的29个核心靶点;KEGG通路结果显示参与癌症途径、PI3K-Akt信号通路等116条KEGG通路;分子对接结果显示二氢血根碱与PTPN11、CSK、NOS2等蛋白结合稳定。结论:多刺绿绒蒿中二氢血根碱含量相对较高,通过网络药理学和分子对接分析得出其通过作用于多靶点、多途径发挥抗氧化的作用。

Objective:Determination of dihydrosanguinarine in Meconopsis horridula and exploration of its antioxidant activity,and the mechanism of dihydrosanguinarine in the treatment of Alzheimer's disease(AD),thyroid-associated ophthalmopathy(TAO),radiation-induced lung injury(RILI),Parkinson's disease(PD)and bronchopulmonary dysplasia(BPD)through its anti-oxidant effect were explored by using network pharmacology and molecular docking technology.Methods:The content of dihydrosanguinarine in M.horridula was determined by HPLC.The anti-oxidant activity was determined by TLC-bioautography assay.Comprehensively use online databases such as Superpre,Genecards and OMIM to collect dihydrosanguinarine targets and targets for treating AD,TAO,RILI,PD and BPD,and build a"compound target disease"and protein interaction network for dihydrosanguinarine treatment of diseases.KEGG pathway enrichment analysis based on the Metascape database and the CytoNCA plug-in.Discovery Studio 2019 Client software was used to conduct molecular docking between the active ingredient and potential targets to verify the binding activity.Results:The results showed that the content of dihydrosanguinarine in Meconopsis horridula was 0.50 mg/g.TLC-bioautography assay results showed that dihydrosanguinarine had anti-oxidant activity.29 core targets of dihydrosanguinarine in the treatment of the above five diseases were obtained through network pharmacological analysis.KEGG pathway results showed that it was involved in 116 KEGG pathways,including cancer pathway and PI3K Akt signaling pathway.Molecular docking results showed that dihydrosanguinarine was stable in binding with PTPN11,CSK,NOS2 and other proteins.Conclusion:Meconopsis horridula had a relatively high content of dihydrosanguinarine.Network pharmacology and molecular docking analyses concluded that it exerts its anti-oxidant effects by acting on multiple targets and pathways.