摘要
目的:采用网络药理学与实验验证研究黄芩素(Baicalein)对癫痫后认知障碍小鼠模型谷氨酸代谢水平的影响,探讨其防治癫痫后认知障碍的作用机制。方法:基于文献检索与网络药理学分析,应用STRING、DAVID在线平台与Cytoscape 3.8.2软件构建蛋白-蛋白互作(PPI)网络,京都基因和基因组百科全书(KEGG)信号通路和基因本体论(GO)功能富集分析。利用分子对接技术预测Baicalein与GRIA2、SLC1A1和SLC1A2基因之间的结合力。体内实验:40只雄性C57BL/6J小鼠随机分为Model组、Baicalein组、VPA组和Baicalein+VPA组,每组10只。给予相应干预后检测小鼠海马组织的炎性浸润情况与SLC1A1、SLC1A2的表达水平变化。体外实验:CHO-GRIA2细胞灌注不同剂量的Baicalein,观察其电流变化。结果:PPI网络中发现兴奋性氨基酸载体1(SLC1A1)、兴奋性氨基酸转运体2(SLC1A2)等40个靶点。KEGG与GO富集分析发现谷氨酸能突触、尼古丁成瘾、离子型谷氨酸受体信号通路、AMPA谷氨酸受体活性、突触后致密膜等与谷氨酸强相关的信号通路。分子对接结果表明,Baicalein与GRIA2、SLC1A1、SLC1A2基因具有较强的结合亲和力。给药后,与Model组比较,Baicalein组与Baicalein+VPA组海马组织炎性浸润率显著降低,各给药组SLC1A1与SLC1A2水平显著升高。CHO-GRIA2灌注1、5、10、20、50μmol/L不同剂量的Baicalein后,其抑制作用逐渐增强,IC50为9.353μmol/L。结论:Baicalein可通过上调谷氨酸转运体SLC1A1和SLC1A2的表达,降低GRIA2受体的表达,改善癫痫后认知障碍的海马炎性浸润情况。
Objective:To study the effects of baicalein on glutamate metabolism in mice with postepileptic cognitive impairment through network pharmacology and experimental verification,and to explore the mechanism of its prevention and treatment of postepileptic cognitive impairment.Methods:Based on literature search and web pharmacological analysis,protein-protein interaction(PPI)network,Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway and gene ontology(GO)functional concentration analysis were constructed by using STRING,DAVID online platform and Cytoscape 3.8.2 software.Molecular docking techniques were used to predict the binding force between baicalein and GRIA2,SLC1A1 and SLC1A2 genes.In the in vivo experiments,40 male mice of C57 were randomly divided into model group,baicalein group,valproate(VPA)group and baicalein+VPA group,with 10 mice in each group.The inflammatory infiltration and the expression levels of SLC1A1 and SLC1A2 were detected after the corresponding intervention.In the in vitro experiments,CHO-GRIA2 cells were perfused with different doses of baicalein to observe the current changes.Results:Forty targets were identified in the PPI network,including excitatory amino acid carrier 1(SLC1A1),excitatory amino acid transporter 2(SLC1A2).KEGG and GO enrichment analyses revealed that glutamatergic synapses,nicotine addiction,ionotropic glutamate receptor signaling pathway,AMPA glutamate receptor activity,postsynaptic dense membrane and other signaling pathways strongly associated with glutamate.Molecular docking results showed that baicalein has strong binding affinity to GRIA2,SLC1A1,SLC1A2 genes.After drug administration,the rate of inflammatory infiltration in hippocampal tissue was significantly reduced in baicalein group and baicalein+VPA group compared with the model group.The level of SLC1A1 and SLC1A2 were significantly increased in all drug administration groups.The inhibitory effect of baicalein on CHO-GRIA2 gradually increased after infusion of different doses of baicalein at 1,5,10,20,and 50μmol/L,with an IC50 of 9.353μmol/L.Conclusions:Baicalein can improve the inflammatory infiltration in the hippocampus of post-epileptic cognitive impairment by up-regulating the expression of the glutamate transporters SLC1A1 and SLC1A2.
作者
李晨阳
赵姣娇
郭晗
陈思软
郝雪南
张炜
LI Chenyang;ZHAO Jiaojiao;GUO Han;CHEN Siruan;HAO Xuenan;ZHANG Wei(Hebei Medical University,Shijiazhuang 050017,China)
出处
《中医药信息》
2024年第6期1-8,18,共9页
Information on Traditional Chinese Medicine
基金
国家自然科学基金项目(81872848)
河北省自然科学基金项目(H2022206211)。
关键词
网络药理学
黄芩素
谷氨酸
癫痫后认知障碍
谷氨酸转运体
Network pharmacology
Baicalein
Glutamate
Post-epileptic cognitive impairment
Glutamate transporter
