基于“肠脑轴”理论探索儿童孤独症伴胃肠道症状的生物学途径

Analysis of biological pathways for children with autism and gastrointestinal symptoms based on the theory of“gut-brain axis”

目的基于“肠脑轴”理论,运用基因芯片的数据挖掘及生物信息学分析初步挖掘孤独症伴胃肠道症状(ASD-GI)相关差异基因(DEGs),探讨其发病机制。方法从基因表达数据库下载芯片数据,筛选差异表达基因进行基因本体论(GO)及京都基因与基因组百科全书数据库(KEGG)分析,构建蛋白质-蛋白质相互作用关系网络(PPI)及运用Cytoscape软件得到核心基因,最后采用分子对接技术将常用治疗药物与核心基因进行对接。结果筛选出共同差异基因20个,其通路富集结果主要包括:PI3K-Akt、Cytokine-cytokine receptor interaction等信号通路。筛选得到10个关键核心基因:CXCL5、IL1B、PIK3CG等。分子对接结果显示血清和糖皮质激素调节激酶1(SGK1)与利培酮结合力很强,人磷酸肌醇-3-激酶催化亚基G肽(PIK3CG)均与利培酮、阿立哌唑、布美他尼具有较好的结合力。结论本研究初步确定ASD-GI发病的关键靶点和途径,在“肠脑轴”理论指导下为ASD-GI的精准治疗提供依据。

Objective Autism spectrum disorder(ASD),referred to as autism,is a neurodevelopmental disorder with social communication disorders,narrow interests or activities,and repetitive stereotyped behaviors as its core symptoms.In addition,patients often have clinical manifestations such as gastrointestinal dysfunction,autistic enterocolitis,anxiety,hyperactivity,etc.Therefore,an in-depth understanding of the mechanism leading to gut-brain communication impairment may help to better understand its underlying pathophysiology.Based on the“gut-brain axis”theory,this study aims to preliminarily mine autism with gastrointestinal symptoms(Gastrointestinal Symptoms in ASD,ASD-GI)related differential genes(DEGs)and to explore its pathogenesis through gene chip data mining and bioinformatics analysis.Methods Using“autism spectrum disorder”as the key words,the GEO database of National Center for Biotechnology Information(NCBI)platform was used to screen and download the microarray data,and the R language clusterProfiler package was used to perform Gene Ontology(GO)and Kyoto Gene Analysis on DEGs.The Encyclopedia of Genomes database(KEGG)was analyzed;the protein-protein interaction network(PPI)was constructed,and the core genes in the protein interaction network were screened by Cytoscape software.Finally,the commonly used therapeutic drugs were docked with the core genes by molecular docking technology.Results The microarray data that met the requirements was GSE87847.The differential expressions of DEGs in peripheral blood and gastrointestinal tissue of ASD-GI and typical developing gastrointestinal symptoms(TD-GI)were compared:257 DEGs in peripheral blood tissue,including 148 up-regulated genes and 109 down-regulated genes;424 DEGs in gastrointestinal tissue,including 160 up-regulated genes and 264 down-regulated genes.By comparing the results of DEGs in peripheral blood and gastrointestinal tissue,20 common differential genes were screened out,and these differential genes were subjected to GO analysis.It was found that biological processes,cellular components,and molecular functions were directly related to immune inflammation and oxidative stress processes,revealing that ASD patients generally had immune disorders.The results of KEGG pathway enrichment analysis mainly included PI3K-Akt signaling pathway,NOD-like receptor signaling pathway,PPAR signaling pathway,Cytokine-cytokine receptor interaction and other signaling pathways.Import 20 common differential genes into the STRING website to construct PPI,and use Cytoscape 3.8.2 software to visualize the hub genes in the network diagram to get 10 key core genes,CXCL5,IL1B,IL7R,FCER1A,EP300,CD36,PTMA,SPI,SGK1,PIK3CG.Molecular docking of key targets with active ingredients Risperidone,Aripiprazole and Bumetanide showed that serum and glucocorticoid-regulated kinase 1(SGK1)had strong binding to Risperidone,and the catalytic subunit G of human phosphoinositide-3-kinase peptides(PIK3CG)have good binding ability to Risperidone,Aripiprazole and Bumetanid.Conclusion This study preliminarily identified the key targets and pathways of ASD-GI pathogenesis,and provided a basis for the precise treatment of ASD-GI under the guidance of the“gut-brain axis”theory.The PI3K/Akt signaling pathway is an important signal transduction pathway related to autism and apoptosis,and the Cytokine-cytokine receptor interaction signaling pathway is an important pathway in the occurrence of ASD-GI,which is related to neuroimmunity and blood-brain barrier damage.Risperidone and aripiprazole may have therapeutic effects by affecting the PI3K-Akt/Wnt signaling pathway.The function of gene-drug docking sites needs to be further designed with basic experiments for mechanism research.The research and development prospects can be based on gut-brain interaction from neuroimaging and other aspects.The correlation of brain network connectivity,the application of acupuncture and traditional Chinese medicine can also provide a research direction for the follow-up rehabilitation treatment of ASD-GI with integrated traditional Chinese and Western medicine.