Yang Xin, He Jia-Yi, Guo Xiang-Xin, Tian Rong
School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China.
Zhongguo Zhong Yao Za Zhi. 2023 May;48(9):2512-2521. doi: 10.19540/j.cnki.cjcmm.20230117.702.
This study aimed to demonstrate the effect of Banxia Baizhu Tianma Decoction(BBTD) on realizing withdrawal of anti-epileptic 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 differen-tial 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 circRNA, 5 miRNA, and 2 mRNA, and a lncRNA-miRNA-mRNA ternary network of 10 lncRNA, 5 miRNA, and 2 mRNA were constructed in ADWG vs BADIG. In conclusion, BBTD can effectively achieve the withdrawal of antiepileptic drugs, which may be related to the transcriptomic regulation of amino acid metabolism.
本研究旨在通过氯化锂-匹罗卡品诱导的癫痫大鼠模型，证明半夏白术天麻汤（BBTD）在实现撤停抗癫痫药物方面的作用，并通过转录组分析探索BBTD与氨基酸代谢之间的关系。将癫痫大鼠分为对照组（Ctrl）、癫痫组（Ep）、BBTD与抗癫痫药物联合组（BADIG）和抗癫痫药物撤停组（ADWG）。Ctrl组和Ep组给予超纯水灌胃12周。BADIG组给予BBTD提取物和卡马西平溶液灌胃12周。ADWG组前6周给予卡马西平溶液和BBTD提取物灌胃，后6周仅给予BBTD提取物灌胃。通过行为观察、脑电图（EEG）和海马神经元形态变化评估治疗效果。采用高通量测序获取海马中与氨基酸代谢相关的差异基因，并通过实时定量聚合酶链反应（RT-qPCR）验证各组海马中的mRNA表达。通过蛋白质-蛋白质相互作用（PPI）网络筛选出枢纽基因，并进行基因本体论（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析。构建了ADWG组与BADIG组的circRNA-miRNA-mRNA和lncRNA-miRNA-mRNA两个ceRNA网络。实验结果表明，与Ep组相比，ADWG组大鼠在行为观察、EEG和海马神经元损伤方面有显著改善。通过转录组分析获得34个与氨基酸代谢相关的差异基因，测序结果经RT-qPCR证实。通过PPI网络获得8个枢纽基因，涉及多个与氨基酸代谢相关的生物学过程、分子功能和信号通路。最后，构建了ADWG组与BADIG组的17个circRNA、5个miRNA和2个mRNA的circRNA-miRNA-mRNA三元转录网络以及10个lncRNA、5个miRNA和2个mRNA的lncRNA-miRNA-mRNA三元网络。综上所述，BBTD能有效实现抗癫痫药物的撤停，这可能与氨基酸代谢的转录组调控有关。
