中国中西医结合杂志2025，Vol.45Issue(11)：1356-1364,9.DOI:10.7661/j.cjim.20250528.015
半夏白术天麻汤调控神经元凋亡lncRNA-miRNA-mRNA转录网络抑制癫痫
Banxia Baizhu Tianma Decoction Inhibits Epilepsy by Improving Neuronal Apoptosis via Regulating lncRNA-miRNA-mRNA Networks
摘要
Abstract
Objective To explore the mechanism underlying the inhibition of neuronal apoptosis in epilepsy by combining Banxia Baizhu Tianma Decoction(BBTD)with carbamazepine(CBZ),and to explore the changes in the lncRNA-miRNA-mRNA network from the perspective of transcriptomics.Methods A lithium chloride-pilocarpine-induced epilepsy rat model was established,and a blank group,a model group,a CBZ group,and a BBTD combined with CBZ group(BCG)were set up,with 10 animals in each group.The blank group and model group were gavaged with ultrapure water for 12 weeks.The CBZ group was gavaged with CBZ solution(125 mg/kg)for 12 weeks,and BCG was gavaged with BBTD extract(21.875 g/kg)and CBZ solution(125 mg/kg)for 12 weeks.Behavioral changes and electroencephalography(EEG)were observed before and after treatment.HE staining and TUNEL staining were employed to detect neuronal damage and apoptosis in the CA1 region of the hippocampus.Immunofluorescence staining was utilized to assess the expression levels of Caspase3,Bcl-2,and Bax proteins.RNA sequencing(RNA-seq)was employed to obtain differential gene expression(DEG)profiles.The model group vs.blank group and BCG vs.model group intersections were utilized to identify common target gene mRNAs for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and to screen apoptotic genes in the KEGG term,thereby constructing the ternary transcriptional network of lncRNA-miRNA-mRNA and protein-protein interaction(PPI)network.Neuronal apoptosis core genes were validated by qRT-PCR.Results Compared with the blank group,the model group exhibited significant neuronal cell damage in histopathological morphology,along with increased seizure frequency,duration,and scores(P＜0.05).The hippocampal CA1 region showed a markedly elevated neuronal apoptosis rate(P＜0.001),accompanied by decreased protein expression levels of Caspase3(P＜0.05)and Bcl-2(P＜0.01),while Bax protein expression was significantly upregulated(P＜0.001).In comparison with the model group,BCG demonstrated alleviated neuronal damage in histopathological examination,reduced seizure frequency,duration,and scores(P＜0.05),along with a significantly decreased apoptosis rate in the hippocampal CA1 region(P＜0.001).The protein expression levels of both Caspase3 and Bcl-2 were reduced(P＜0.05),while Bax protein expression remained significantly elevated(P＜0.001).Transcriptome sequencing results showed that Model group vs.Blank group and BCG vs.Model group intersection 344 mRNAs of common target genes were obtained.Totally 344 mRNAs were found to be related to neuronal apoptosis revealed by the GO enrichment analysis and KEGG pathway analysis.A total of 31 apoptotic genes were screened from KEGG apoptotic pathway entries.The core ceRNA regulatory network involved in neuronal apoptosis was found to consist of six mRNAs(Ntrk1,Htr6,Ptgs2,Perp,Rasgrp3,Tacr2),four miRNAs(rno-miR-409b,rno-miR-433-3p,rno-miR-382-3p,rno-miR-3592),and 11 lncRNAs.Conclusion The combination of BBTD with CBZ attenuate and ameliorate neuronal damage and inhibit neuronal apoptosis,exerting a significant antiepileptic effect via regulating lncRNA-miRNA-mRNA networks.关键词
半夏白术天麻汤/卡马西平/细胞凋亡/lncRNA-miRNA-mRNA三元转录网络/癫痫/转录组学分析/中药复方/中西医结合Key words
Banxia Baizhu Tianma Decoction/Carbamazepine/apoptosis/ternary transcription network of lncRNA-miRNA-mRNA/epilepsy/transcriptomic analysis/Chinese herbal compound/integrative medicine引用本文复制引用
郭向鑫,杨欣,何佳忆,开胤桦,朱兴丹,蒋萃,田茸..半夏白术天麻汤调控神经元凋亡lncRNA-miRNA-mRNA转录网络抑制癫痫[J].中国中西医结合杂志,2025,45(11):1356-1364,9.基金项目
国家自然科学基金资助项目(No.81973929) （No.81973929）
全国中医药创新骨干人才培训项目(No.国中医药办人教函[2019]91号) （No.国中医药办人教函[2019]91号）
四川省自然科学基金资助项目(No.2023NSFSC18) （No.2023NSFSC18）
成都中医药大学杏林人才苗圃项目(No.MPRC2022003) （No.MPRC2022003）
