基于转录组学分析探究半夏白术天麻汤撤药治疗的氨基酸代谢特征  

Amino acid metabolism characteristics of Banxia Baizhu Tianma Decoction in realizing drug withdrawal based on transcriptomic analysis

作　　者：杨欣 何佳忆 郭向鑫 田茸[1] YANG Xin;HE Jia-yi;GUO Xiang-xin;TIAN Rong(School of Basic Medical Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学基础医学院,四川成都611137

出　　处：《中国中药杂志》2023年第9期2512-2521,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81973929);全国中医药创新骨干人才培训项目(国中医药办人教函〔2019〕91号)。

摘　　要：该研究基于转录组学分析方法,结合氯化锂-匹罗卡品诱导的癫痫大鼠模型,明确半夏白术天麻汤撤药治疗方案的有效性,探究半夏白术天麻汤撤药治疗与氨基酸代谢的关系。构建氯化锂-匹罗卡品癫痫大鼠模型,设置对照组(control group,Ctrl)、模型组(epilepsy group,Ep)、中西药结合治疗组(Banxia Baizhu Tianma Decoction&antiepileptic drug integrative group,BADIG)、撤药治疗组(antiepileptic drug withdrawal group,ADWG)。Ctrl和Ep以超纯水灌胃12周;BADIG灌胃半夏白术天麻汤浸膏和卡马西平溶液12周;ADWG前6周灌胃半夏白术天麻汤浸膏和卡马西平溶液,后6周仅灌胃半夏白术天麻汤浸膏。通过大鼠行为学、脑电图和神经元细胞病理变化评估疗效。高通量测序获取海马组织中氨基酸代谢相关差异基因,采用qRT-PCR验证各组海马组织中mRNA的表达,构建蛋白-蛋白互作(protein-protein interaction,PPI)网络筛选核心基因,并对其进行基因本体(Gene Ontology,GO)功能和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。针对ADWG vs BADIG分别构建circRNA-miRNA-mRNA和lncRNA-miRNA-mRNA 2组ceRNA网络。结果显示,与Ep比较,ADWG大鼠在行为学、脑电图以及神经元病理形态学上均得到显著改善。转录组学分析共筛选出34个氨基酸代谢相关基因,并通过qRT-PCR实验对测序结果进行验证,构建PPI网络获取8个核心基因,涉及多条氨基酸代谢相关生物学过程、分子功能以及信号通路。根据氨基酸代谢相关基因,在ADWG vs BADIG中构建了由17个circRNA、5个miRNA和2个mRNA构成的circRNA-miRNA-mRNA三元转录网络,以及由10个lncRNA、5个miRNA和2个mRNA构成的lncRNA-miRNAmRNA三元转录网络。综上所述,半夏白术天麻汤能有效实现抗癫痫药物的撤药,其机制可能与氨基酸代谢的转录组学调控有关。This study aimed to demonstrate the effect of Banxia Baizhu Tianma Decoction(BBTD)on realizing withdrawal of antiepileptic drugs and explore the relationship between BBTD and the amino acid metabolism by transcriptomic analysis in the rat model of epilepsy induced by lithium chloride-pilocarpine.The rats with epilepsy were divided into a control group(Ctrl),an epilepsy group(Ep),a BBTD&antiepileptic drug integrative group(BADIG),and an antiepileptic drug withdrawal group(ADWG).The Ctrl and Ep were given ultrapure water by gavage for 12 weeks.The BADIG was given BBTD extract and carbamazepine solution by gavage for 12 weeks.The ADWG was given carbamazepine solution and BBTD extract by gavage for the former 6 weeks,and then only given BBTD extract for the latter 6 weeks.The therapeutic effect was evaluated by behavioral observation,electroencephalogram(EEG),and hippocampal neuronal morphological changes.High-throughput sequencing was used to obtain amino acid metabolism-related differential genes in the hippocampus,and the mRNA expression in the hippocampus of each group was verified by real-time quantitative polymerase chain reaction(RT-qPCR).The hub genes were screened out through protein-protein interaction(PPI)network,and Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed.Two ceRNA networks,namely circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA,were constructed for ADWG vs BADIG.The experimental results showed that compared with those in Ep,rats in ADWG were significantly improved in the behavioral observation,EEG,and hippocampal neuronal impairment.Thirty-four amino acid metabolism-related differential genes were obtained by transcriptomic analysis,and the sequencing results were confirmed by RT-qPCR.Eight hub genes were obtained through PPI network,involving several biological processes,molecular functions,and signal pathways related to amino acid metabolism.Finally,the circRNA-miRNA-mRNA ternary transcription network of 17 circ

关 键 词：抗癫痫药物撤药 氨基酸代谢 半夏白术天麻汤 海马 癫痫 转录组学分析 

分 类 号：R285.5[医药卫生—中药学]