基于网络药理学及实验验证探讨飞龙掌血治疗缺血性脑卒中的作用机制  

Exploration of the Mechanism of T oddalia asiatica in the Treatment of Ischemic Stroke:Based on Network Pharmacology and Experimental Validation

作　　者：高建红 杨丹 王刚 宋天英 赵方毓 陈显兵 GAO Jian-hong;YANG Dan;WANG Gang;SONG Tian-ying;ZHAO Fang-yu;CHEN Xian-bing(Hubei Key Laboratory of Rheumatology and Intervention in Rheumatic Diseases,Hubei Minzu Universit,Hubei Minzu University,Enshi Hubei 445000,Chinay;Hubei Clinical Medicine Research Center of Kidney Diseases;Health Science Center,Hubei Minzu University,Enshi Hubei 445000,China)

机构地区：[1]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [2]湖北省肾脏病临床医学研究中心 [3]湖北民族大学医学部,湖北恩施445000

出　　处：《中国药理学通报》2024年第7期1375-1383,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金项目(No 82260821);恩施州科技局项目(No JCY2021000054;2018-15);湖北民族大学博士启动基金项目(No MD2020B014);湖北民族大学研究生教育创新项目(No MYK2023069)。

摘　　要：目的基于网络药理学、分子对接技术以及动物实验,阐释飞龙掌血治疗缺血性脑卒中(ischemic stroke,IS)的作用机制。方法利用数据库筛选飞龙掌血化学成分和IS靶点,构建PPI网络,建立KEGG通路富集分析,进行成分与靶点的分子对接,最后通过动物实验,验证药物对PI3K/AKT/mTOR通路及自噬的影响。动物实验中,制备MCAO大鼠模型,随机分为模型组、盐酸多奈哌齐组、飞龙掌血组,并设假手术组,观察药物对大鼠海马区、皮质区神经元的病理变化情况,免疫组化检测mTOR表达及定位,Western blot检测PI3K、p-PI3K、AKT、p-AKT、mTOR以及自噬标识物LC3-Ⅱ、p62表达情况。结果最终筛选到22个活性成分,AKT1、MAPK3等29个核心靶点,PI3K/AKT、MAPK等194条信号通路。活性成分与IS相关靶点的结合稳定。动物实验结果表明,与假手术组相比,模型组大鼠海马区、皮质区神经元分布稀疏紊乱,尼氏小体数量减少,胞质空泡化,海马区、皮质区mTOR阳性细胞表达减少,海马组织p-PI3K、p-AKT、mTOR和p62表达降低(P<0.05,P<0.01),LC3-Ⅱ表达升高(P<0.01)。与模型组相比,飞龙掌血组和盐酸多奈哌齐组大鼠有效改善上述指标。结论通过网络药理学分析发现,飞龙掌血具有多成分、多靶点、多通路治疗IS的潜力。动物实验表明,飞龙掌血可以保护海马区、皮质区神经元结构,这可能与抑制PI3K/AKT/mTOR通路介导的过度自噬有关。Aim This study aims to investigate the therapeutic effect and underlying mechanism of Toddalia asiatica in the treatment of ischemic stroke(IS),utilizing network pharmacology,molecular docking technology,and animal experiments.Methods To screen the chemical components of Toddalia asiatica and its targets related to IS,a database was utilized.A protein-protein interaction(PPI)network was constructed,followed by KEGG pathway enrichment analysis.Molecular docking was performed to investigate the interaction between the components and target proteins.Finally,the effects of the drug on the PI3K/AKT/mTOR pathway and autophagy were validated through animal experiments.We established a middle cerebral artery occlusion(MCAO)rat model and divided the rats into the model group,Donepezil hydrochloride group,Toddalia asiatica group,and sham operation group randomly.Observed the pathological changes in neurons of the rat hippocampal and cortical regions induced by the drug,performed immunohisto-chemical analysis to detect and localize mTOR expression,and used Western blot to assess the expression levels of PI3K,p-PI3K,AKT,p-AKT,mTOR,as well as autophagy markers(LC3-Ⅱand p62).Results A total of 22 active ingredients from Toddalia asiatica,including AKT1 and MAPK3,were identified through screening.Additionally,194 signaling pathways,such as PI3K/AKT and MAPK,were analyzed.The active compounds in Toddalia asiatica demonstrated stable binding affinity with targets associated with ischemic stroke.The results of the animal experiment indicated that,compared to the sham-operated group,the neuronal distribution in the hippocampal and cortical regions of the model group rats became sparser and more disorganized.There was a decrease in the number of Nissl bodies and cytoplasmic vacuolization.The expression of mTOR-positive cells in the hippocampal and cortical regions was reduced.Additionally,the expression levels of p-PI3K,p-AKT,mTOR,and p62 in the rat hippocampal tissue decreased(P<0.05,P<0.01),while the expression of LC3-Ⅱincreas

关 键 词：网络药理学 飞龙掌血 缺血性脑卒中 PI3K/AKT/mTOR信号通路 自噬 实验 

分 类 号：R-332[医药卫生] R285.5R322.81R319R743.31