中国药科大学学报2024，Vol.55Issue(4)：512-521,10.DOI:10.11665/j.issn.1000-5048.2023111303
基于全长转录组分析探讨熊果酸抗癫痫和神经保护的机制
Antiepileptic and neuroprotective mechanism of ursolic acid based on full-length transcriptome analysis
摘要
Abstract
This study explores the potential antiepileptic mechanism of ursolic acid(UA)and its improvement of GABAergic interneuron damage induced by epilepsy based on transcriptome analysis.Hippocampal tissues from rats in the control group(NC group),epilepsy group(SE group),and epilepsy UA treatment group(UA group)were subjected to full-length transcriptome sequencing.The obtained sequencing data were analyzed,using gene ontology(GO),the Kyoto Encyclopedia of Genes and Genomes(KEGG),and protein-protein interaction(PPI)to perform the analysis of differential genes(DEGs).The expression levels of key differential genes were verified using RT-qPCR in hippocampal tissue.Finally,an epilepsy in vitro model was constructed on primary neurons,RT-qPCR was used to verify the expression levels of key differential genes,and the expression level of GABAA receptor γ2 subunit(GABRG2)on neurons was further examined using immunofluorescence and Western blot.The heatmap of pairwise sample expression correlation and the clustering analysis of differentially expressed genes showed that the SE group was farthest from the NC group,and that after UA treatment,the overall trend shifted towards the normal group.Compared with the SE group,a total of 220 differential genes were screened in the UA group,including 143 upregulated genes and 77 downregulated genes.GO enrichment analysis showed that it involved three processes in the primary classification:biological processes,cellular components,and molecular functions.KEGG pathway enrichment analysis showed that DEGs were involved in 36 biological pathways,including cAMP signaling pathway and calcium signaling pathway.PPI analysis showed that DEGs were closely related to GABA and inflammation.RT-qPCR results showed that UA treatment increased the expression levels of GABA receptor-related gene(Gng4),GABA synthesis-related gene(Camk2a,Vgf,and Npy)and inflammation-related gene(Timp1 and Spp1)in hippocampal tissue,and decreased the expression levels of GABA synthesis-related gene(Nptx2)and cAMP-related pathway gene(Gnas).It further confirmed that UA treatment increased the expression levels of Gng4 and Camk2a on neurons and decreased the expression level of Gnas.Immunofluorescence and Western blot results showed that,compared with the SE group,the expression level of GABRG2 on primary neurons increased after UA treatment.This study enriched the transcriptome data of UA's antiepileptic effect and laid a theoretical foundation for further research on UA's antiepileptic and neuroprotective effects.关键词
熊果酸/癫痫/全长转录组/测序分析/GABAKey words
ursolic acid/epilepsy/full-length transcriptome/sequencing analysis/GABA分类
医药卫生引用本文复制引用
李娟娟,黄越,王雅禾,张莲香,强媛媛,郭乐,刘昆梅..基于全长转录组分析探讨熊果酸抗癫痫和神经保护的机制[J].中国药科大学学报,2024,55(4):512-521,10.基金项目
国家自然科学基金项目(No.82360711,No.32070930) （No.82360711,No.32070930）
宁夏自然科学基金项目(No.2023AAC03155) （No.2023AAC03155）
宁夏颅脑疾病重点实验室主任基金项目(No.LNZR202304) （No.LNZR202304）
宁夏回族自治区青年拔尖人才项目 This study was supported by the National Natural Science Foundation of China(No.82360711,No.32070930) （No.82360711,No.32070930）
the Natural Science Foundation of Ningxia(No.2023AAC03155) （No.2023AAC03155）
Ningxia Craniocerebral Diseases Key Laboratory Director Fund Project(No.LNZR202304) （No.LNZR202304）
and Ningxia Youth Top Talent Project （）
