基于动物实验和网络药理学研究艾纳香对急性肝损伤保护作用机制

Study on the protective mechanism of blumea balsamifera on acute liver injury based on animal experiment and network pharmacology

关键词:  艾纳香  急性肝损伤  网络药理学  四氯化碳  机制

Key words:  Blumea balsamina  acute liver injury  network pharmacology  CCl₄  mechanism

摘要:

[目的]基于动物实验和网络药理学的方法研究艾纳香对于急性肝损伤（ALI）的保护作用及潜在机制。[方法]腹腔注射CCl4构建小鼠急性肝损伤动物模型，通过观察小鼠血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）水平变化，初步探究艾纳香对急性肝损伤的保护作用。采用网络药理学方法检索艾纳香活性成分，预测作用靶点，构建“药物-活性成分-疾病-靶点”网络、蛋白质-蛋白质相互作用（PPI）网络，利用分子对接方法对所得化合物和靶点进行对接验证，通过基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析，进一步探究艾纳香治疗ALI的作用机制。[结果]实验结果证明艾纳香可明显缓解CCl4诱导的急性肝损伤，降低小鼠血清ALT、AST水平，差异具有统计学意义（P＜0.05）；网络药理学分析获得艾纳香的主要活性成分36个，关键成分10个，包括羟基芫花素、木犀草素、山奈酚、芫花素、华良姜素、槲皮素-3，3'-二甲醚、金圣草素、香叶木素、圣草酚、5，3'-二羟基-7，4'-二甲氧基黄酮；艾纳香作用靶点171个，筛选核心靶点10个，包括丝氨酸/苏氨酸激酶（AKT1）、血管内皮生长因子A（VEGFA）、表皮生长因子受体（EGFR）、雌激素受体（ESR1）、半胱氨酸蛋白酶（CASP3）、白细胞介素-1β（IL-1β）、热休克蛋白90α家族A级成员1（HSP90AA1）、缺氧诱导因子1 （HIF1A）、丝裂原活化蛋白激酶3（MAPK3）、原癌基因酪氨酸激酶（SRC）；GO分析显示生物过程主要富集在蛋白磷酸化、氮化合物的细胞反应、细胞迁移的正向调节，KEGG富集涉及癌症通路、PI3K-Akt信号通路、癌症中的微小RNA、脂质和动脉粥样硬化、MAPK信号通路等通路。[结论]研究初步证实艾纳香提取物对CCl4所致急性肝损伤产生显著的保护作用，其可能是通过调控AKT1、VEGFA等核心靶点及癌症、PI3K-Akt等信号通路，减轻炎症反应、抑制氧化应激，从而产生肝保护作用。

Abstract:

[Objective] This study was based on animal experiments and network pharmacology tostudy the protective effect and potential mechanism of Blumea balsamifera on acute liver injury(ALI).[Methods] Acute liver injury model was induced by intraperitoneal injection of CCl₄ in mice. The protective effect of Blumea balsamifera on acute liver injury was preliminarily explored by observing the changes of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in mice. The network pharmacology method was used to retrieve the active components of Blumea balsamina,predict the action target,construct the "Drug-active component-target-disease" network and protein-protein Interaction Network(PPI),verify the docking between the obtained compounds and the targets by molecular docking method,and further explore the action mechanism of Blumea balsamina in the treatment of ALI by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrich-ment analysis.[Results] The results showed that Blumea balsamina could significantly alleviate the acute liver injury induced by CCl₄ and significantly reduce the levels of ALT and AST in mice(P＜0.05);The network pharmacology analysis showed that there were 36 main active componentsand 10 key components,including3-hydroxygenkwanin,luteolin,kaempferol,5,4'dihydroxy-7-methoxyflavone,Kumatakenin,Quercetin3,3'-dimethyl etheh,Chrysoeriol,Diosmetin,Eriodictyol, Pilloin. There were 171 targets for the action of Blumea balsamina,and 10 core targets were screened,including RAC-alpha serine/threonine-protein kinase (AKT1),vascular endothelial growth factor A (VEGFA),Epidermal growth factor receptor(EGFR),Estrogen receptor(ESR1),Caspase-3(CASP3),Interleukin-1 beta(IL1B),heat shock protein 90α family class A member1(HSP90AA1),Hypoxia-inducible factor 1-alpha(HIF1A),Mitogen-activated protein kinase 3(MAPK3),Protooncogene tyrosine-protein kinase Src(SRC). The results of moleculardocking showed that the key components and core targets screened bynetwork pharmacology could bind spontaneously. Go analysis shows that biological processes are mainly concentrated in protein phosphorylation,cellular response to nitrogen compound,positive regulation of cell migration,regulation of kinase activity. KEGG enrichment involves cancer pathway,PI3K-Akt signaling pathway,microRNA in cancer,lipid and atherosclerosis,MAPK signaling pathway and so on.[Conclusion]This study preliminarily confirmed that the extract of Blumeabalsamina has a significant protective effect on CCl₄ induced acute liver injury,which may be through regulating AKT1,VEGFA and other core targets and cancer,PI3K-Akt and other signal path-ways,reducing inflammatory response and inhibiting oxidative stress,so as to produce liver protective effect.