半夏白术天麻汤对癫痫大鼠海马miRNA—146a—5p表达的影响及生物信息学分析(1)
摘要：目的 通過对癫痫大鼠海马miRNA-146a-5p进行靶基因预测及信号通路生物信息学分析，为探讨癫痫的可能发病机制及半夏白术天麻汤抗癫痫作用机制的研究奠定基础。方法 采用氯化锂-匹鲁卡品诱导制作癫痫发作大鼠模型。将实验大鼠随机分为正常组、模型组、中药组，每组20只。使用miRNA芯片技术分析检测大鼠海马神经元细胞miRNA表达谱。实时荧光定量PCR检测miRNA-146a-5p表达。利用miRDB对miRNA-146a-5p进行靶基因预测，并运用miRTarBase、DAVID进行GO功能和KEGG信号通路的富集分析。结果 行为学观察显示，中药组大鼠发作次数和级别均较模型组明显降低。miRNA芯片结果显示，模型组miRNA-146a-5p表达水平是正常组的2.107倍(P<0.05)，经半夏白术天麻汤治疗后其表达量降至1.377倍(P<0.05)。RT-PCR结果与芯片结果一致，表达差异有统计学意义(P<0.05)。miRNA-146a-5p靶基因预测结果共有140个靶基因，GO注释共得到14个GO生物学过程注释信息(P<0.05)，9个细胞组分注释信息(P<0.05)，11个分子功能注释信息(P<0.05)。KEGG生物通路富集显示，其140个靶基因显著富集于EB病毒感染信号通路，以及甲状腺激素信号通路上(P<0.05)。结论 miRNA-146a-5p与癫痫发生后的炎症反应密切相关，半夏白术天麻汤可能通过控制癫痫发生后的炎症反应发挥抗癫痫的效应。
关键词：癫痫；海马；miRNA-146a-5p；半夏白术天麻汤；大鼠
DOI：10.3969/j.issn.1005-5304.2018.07.009
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2018)07-0034-07
Abstract： Objective To lay the foundation for studying the possible pathogenesis of epilepsy and the anti-epileptic mechanism of Banxia Baizhu Tianma Decoction through the bioinformatic analysis of target gene prediction and signal pathway of miRNA-146a-5p in hippocampus of epileptic rats. Methods Lithium-pilocarpine was used to induce seizures in rat models. The experiment rats were randomly divided into normal control group， model group， Banxia Baizhu Tianma Decoction group， with 20 rats in each group. The method of miRNA expression profiling was used to observe the miRNA differential expression of hippocampus neuron cell of rats. The expression level of miRNA-146a-5p was detected by real-time quantitative PCR. MiRDB was used for target gene prediction of miRNA-146a-5p， and miRTarBase and DAVID were used for enrichment analysis on the GO function and KEGG signaling pathway. Results The attack times and grades of the rats in Banxia Baizhu Tianma Decoction group were significantly lower than those in the model group from behavioral observation. MiRNA microarray analysis showed that the expression level of miRNA-146a-5p in model group was 2.107 times normal control group (P<0.05)， and the expression level decreased to 1.377 times after treatment with Banxia Baizhu Tianma Decoction (P<0.05). The results
of RT-PCR was consistent with that of miRNA microarray， with statistical significance (P<0.05). MiRNA-146a-5p target gene prediction results had 140 target genes by GO， and there were 14 annotation information of biological process (P<0.05)， 9 annotation information of cellular component (P<0.05)， 11 annotation information of molecular function (P<0.05). Enrichment analysis of KEGG biological pathway showed that 140 target genes of miRNA-146a-5p were enriched in EB virus infection signal pathway and thyroid hormone signaling pathway (P<0.05). Conclusion miRNA-146a-5p is closely related to the inflammatory reaction after epilepsy， and Banxia Baizhu Tianma Decoction can control epilepsy possibly by controlling the inflammatory reaction after epilepsy., http://www.100md.com(田茸 舍雅莉 董晓丽 史正刚 陈丽 程小丽 王宇)
关键词：癫痫；海马；miRNA-146a-5p；半夏白术天麻汤；大鼠
DOI：10.3969/j.issn.1005-5304.2018.07.009
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2018)07-0034-07
Abstract： Objective To lay the foundation for studying the possible pathogenesis of epilepsy and the anti-epileptic mechanism of Banxia Baizhu Tianma Decoction through the bioinformatic analysis of target gene prediction and signal pathway of miRNA-146a-5p in hippocampus of epileptic rats. Methods Lithium-pilocarpine was used to induce seizures in rat models. The experiment rats were randomly divided into normal control group， model group， Banxia Baizhu Tianma Decoction group， with 20 rats in each group. The method of miRNA expression profiling was used to observe the miRNA differential expression of hippocampus neuron cell of rats. The expression level of miRNA-146a-5p was detected by real-time quantitative PCR. MiRDB was used for target gene prediction of miRNA-146a-5p， and miRTarBase and DAVID were used for enrichment analysis on the GO function and KEGG signaling pathway. Results The attack times and grades of the rats in Banxia Baizhu Tianma Decoction group were significantly lower than those in the model group from behavioral observation. MiRNA microarray analysis showed that the expression level of miRNA-146a-5p in model group was 2.107 times normal control group (P<0.05)， and the expression level decreased to 1.377 times after treatment with Banxia Baizhu Tianma Decoction (P<0.05). The results
of RT-PCR was consistent with that of miRNA microarray， with statistical significance (P<0.05). MiRNA-146a-5p target gene prediction results had 140 target genes by GO， and there were 14 annotation information of biological process (P<0.05)， 9 annotation information of cellular component (P<0.05)， 11 annotation information of molecular function (P<0.05). Enrichment analysis of KEGG biological pathway showed that 140 target genes of miRNA-146a-5p were enriched in EB virus infection signal pathway and thyroid hormone signaling pathway (P<0.05). Conclusion miRNA-146a-5p is closely related to the inflammatory reaction after epilepsy， and Banxia Baizhu Tianma Decoction can control epilepsy possibly by controlling the inflammatory reaction after epilepsy., http://www.100md.com(田茸 舍雅莉 董晓丽 史正刚 陈丽 程小丽 王宇)