摘要
目的采用网络药理学和分子对接技术及分子动力学技术,研究骆驼刺治疗脓毒症作用机制,并进行动物实验验证。方法首先筛选骆驼刺有效成分及其作用靶点,同时筛选治疗脓毒症的相关作用靶点,构建蛋白质相互作用(PPI)网络,并进行拓扑学分析,绘制中药-疾病-靶点网络图。其次对网络图中核心靶点进行京都基因与基因组百科全书富集分析,同时进行基因本体功能富集分析;接着对核心靶点进行分子对接和分子动力学模拟实验验证。最后采用小鼠进行动物实验验证。结果筛选出骆驼刺活性成分30个,度值排名前5为(-)-儿茶素酸酯、金雀异黄酮、山奈酚、槲皮素、表没食子儿茶素,作用靶点196个;脓毒症疾病相关靶点2144个,骆驼刺-脓毒症交集靶点105个,核心靶点为TNF、IL-6、AKT1、VEGFA、CASP3、IL-1β等。涉及PI3K-Akt、TNF、HIF-1、AGE-RAGE、IL-17等信号通路,介导炎症反应、细胞凋亡等生物过程,发挥对脓毒症的治疗作用。分子对接结果显示,骆驼刺黄酮类化合物与各关键靶点结合性能均良好,其中结合能最低最稳定的构象为(-)-epigallocatechin gallate-IL-6和quercetin-IL-6,对两对复合物进行分子动力学模拟,结果表明,通过静电、范德华势能和氢键的共同作用,可以达到稳定结合的效果。动物实验证实骆驼刺可抑制脓毒症小鼠肺组织内PI3K/Akt信号通路的激活,降低Caspase-3、VEGF蛋白表达,减少外周血炎症因子TNF-α、IL-1β、IL-6的分泌,减轻组织及脏器炎性损伤。结论骆驼刺对脓毒症的治疗作用是通过多生物过程、多靶点、多通路来实现的。本研究为骆驼刺的临床应用以及脓毒症治疗提供了一定的理论依据。
Objective Network pharmacology and molecular docking and molecular dynamics techniques were used to investigate the mechanism of action of Alhagi sparsifolia Shap.in the treatment of sepsis and to perform animal experimental verification.Methods First,we screened the effective ingredients and their action targets of Alhagi sparsifolia Shap.,meanwhile,screened relevant action targets for the treatment of sepsis,constructed a protein interaction(PPI)network,and performed topology analysis to draw a TCM disease target network diagram.Second,Kyoto Encyclopedia of genes and genomes enrichment analysis was performed for core targets in the network diagram,along with gene ontology functional enrichment analysis.This was followed by molecular docking and molecular dynamics simulation experiment validation of the core targets.Finally,mice were used for the verification of animal experiments.Results Thirty active components of Alhagi sparsifolia Shap.were screened out,and the top 5 ranked by degree value were quercetin,(-)-epigallocatechin,(-)-Epigallocatechin Gallate,genistein,kaempferol and epigallocatechin with 196 action targets;2144 disease-related targets for sepsis,105 targets for Alhagi sparsifolia Shap.-sepsis intersection,and the core targets were TNF,IL-6,AKT1,VEGFA,CASP3,IL-1βEt al.PI3K-Akt,TNF,HIF-1,AGE-RAGE,IL-17 and other signaling pathways are involved to mediate inflammatory responses,apoptosis and other biological processes to exert therapeutic effects on sepsis.Molecular docking results showed that camelina flavanoids bound equally well to each key target,among which the conformations with the lowest binding energy were(-)-Epigallocatechin Gallate-IL-6 and quercetin-IL-6.Molecular dynamics simulations were performed on the two pairs of complexes,and the results indicated that the stable binding could be achieved through a combination of electrostatic,van der Waals potential,and hydrogen bonding interactions.Animal experiments confirmed that Alhagi sparsifolia Shap.could inhibit the activation of PI3K/Akt signaling pathway,decrease the protein expression of Caspase-3,VEGF and reduced peripheral blood inflammatory factors secretion of TNF-α、IL-1β and IL-6,alleviating inflammatory injury in tissues and organs.Conclusion The therapeutic effect of Alhagi sparsifolia Shap.on sepsis is achieved through multi biological processes,multi targets,and multi pathways.It provides a certain theoretical basis for the clinical application of camel spines as well as sepsis treatment.
作者
邹炙臻
邓喜玲
王云来
张杰
董江涛
柳小玲
梁粟
王菊
张辉
吴江东
章乐
吴芳
张万江
Zou Zhizhen;Deng Xiling;Wang Yunlai;Zhang Jie;Dong Jiangtao;Liu Xiaoling;Liang Su;Wang Ju;Zhang Hui;Wu Jiangdong;Zhang Le;Wu Fang;Zhang Wanjiang(Department of Pathophysiology,School of Medicine,Shihezi University/High Incidence of Local and Ethnic Groups in Xinjiang Ministry of Education Key Laboratory,Shihezi 832002,China;Department of Parmacy,School of Parmacy,Shihezi University/Xinjiang Phytomedicine Resource and Utilization Ministry of Education Key Laboratory,Shihezi 832002,China;The Second Affiliated Hospital Zhejiang University School of Medicine,Hangzhou 310000,China;The First Affiliated Hospital of Shihezi University School of Medicine,Shihezi 832003,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第9期3024-3036,共13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地区科学基金项目(82060021):Tim-3调控肺泡巨噬细胞极化和功能对脓毒症急性肺损伤发生发展的影响及其机制研究,负责人:张万江
石河子大学2020年度自主资助支持校级科研立项项目(ZZZC202013A):Blimp-1多靶点调控脓毒症时机体炎症因子表达及分泌的研究,负责人:吴芳。
关键词
骆驼刺
脓毒症
网络药理学
分子对接
分子动力学
作用机制
Alhagi sparsifolia Shap.
Sepsis
Network pharmacology
Molecular docking
Molecular dynamics
Mechanism
