作者：仲婷，景燕，陆明佳，党辉，李红燕

单位：

830000新疆维吾尔自治区乌鲁木齐市，新疆维吾尔自治区人民医院神经内科 新疆脑卒中与神经系统罕见病临床医学研究中心

单位（英文）：

Department of Neurology, People's Hospital of Xinjiang Uygur Autonomous Region/Xinjiang Clinical Medical Center forStroke and Rare Neurological Diseases, Urumqi 830000, China

关键词：

癫痫；特发性癫痫；微RNAs；hsa-let-7b-3p；hsa-miR-92a-1-5p；hsa-miR-6810-3p

关键词（英文）：

Epilepsy; Idiopathic epilepsy; MicroRNAs; hsa-let-7b-3p; hsa-miR-92a-1-5p; hsa-miR-6810-3p

中图分类号：

　R 256.61

DOI：

10.12114/j.issn.1008-5971.2023.00.179

基金项目：

新疆维吾尔自治区自然科学基金资助项目（2019D01C149）

摘要：

目的　分析特发性癫痫患者外周血差异表达miRNA。方法　选取2019年1月至2020年1月在新疆维吾尔自治区人民医院神经内科治疗的特发性癫痫患者12例为病例组，选取同期于新疆维吾尔自治区人民医院门诊进行体检的健康者12例为对照组。采集受试者外周血，采用Illumina HiSeq 2000测序仪进行高通量测序，采用edgeR软件筛选差异表达miRNA，采用qPCR检测差异表达miRNA表达水平。结果　共完成24个样本测序。各样本原始序列数据统计分析结果显示，原始序列中碱基的测序质量值＞20的碱基占总碱基的百分比为92.56%～97.32%，原始序列中碱基的测序质量值＞30的碱基占总碱基的百分比为88.01%～93.50%，碱基G和C的数量总和占总碱基数量的百分比为45.29%～52.09%；各样本样品待分析数据统计分析结果显示，原始序列中碱基的测序质量值＞20的碱基占总碱基的百分比为95.87%～98.85%，原始序列中碱基的测序质量值＞30的碱基占总碱基的百分比为95.30%～96.85%，碱基G和C的数量总和占总碱基数量的百分比为40.38%～47.58%。共检测出146个差异表达miRNA，与对照组相比，研究组共有98个miRNA表达上调，48个miRNA表达下调；其中差异倍数最大的5个差异表达miRNA分别为hsa-let-7b-3p、hsa-let7f-1-3p、hsa-miR-92a-1-5p、hsa-miR-219a-2-3p、hsa-miR-6810-3p。扩增曲线分析结果显示，样本均经历了良好的扩增过程。熔解曲线分析结果显示，大多数曲线为单峰。病例组hsa-let-7b-3p、hsa-miR-92a-1-5p表达水平低于对照组，hsa-miR-6810-3p表达水平高于对照组（P＜0.05）。结论　与健康人相比，特发性癫痫患者外周血共有98个miRNA表达上调，48个miRNA表达下调；其中差异倍数最大的5个差异表达miRNA分别为hsa-let-7b-3p、hsa-let-7f-1-3p、hsa-miR-92a-1-5p、hsa-miR-219a-2-3p、hsa-miR-6810-3p，且可能只有hsa-let-7b-3p、hsa-miR-92a-1-5p、hsa-miR-6810-3p参与了特发性癫痫的发生发展。

英文摘要：

Objective To analyze differentially expressed miRNA in peripheral blood of patients with idiopathicepilepsy. Methods Twelve patients with idiopathic epilepsy who were treated in Department of Neurology, People's Hospital ofXinjiang Uyghur Autonomous Region from January 2019 to January 2020 were selected as the case group. Twelve healthy subjectswho underwent physical examination in the Outpatient Department of People's Hospital of Xinjiang Uyghur Autonomous Regionduring the same period were selected as the control group. The peripheral blood of the subjects was collected, and the highthroughput sequencing was performed using Illumina HiSeq 2000 sequencer. The differentially expressed miRNA was screenedby edgeR software, and the expression levels of the differentially expressed miRNA were detected by qPCR. Results A total of24 samples were sequenced. The statistical analysis results of raw reads of each sample showed that the bases with sequencingquality value > 20 in the original sequence accounted for 92.56%-97.32% of the total bases. The percentage of bases with thesequencing mass value > 30 to the total bases in the original sequence was 88.01%-93.50%, and the percentage of the sum of thenumbers of bases G and C to the total number of bases was 45.29%-52.09%. The statistical analysis results of clean reads of eachsample showed that the bases with sequencing quality value > 20 in the original sequence accounted for 95.87%-98.85% of thetotal bases, the percentage of bases with the sequencing mass value > 30 to the total bases in the original sequence was 95.30%-96.85%, and the percentage of the sum of the numbers of bases G and C to the total number of bases was 40.38%-47.58%. Atotal of 146 differentially expressed miRNAs were detected. Compared with the control group, 98 miRNAs in the study groupwere up-regulated and 48 miRNAs were down-regulated. Among them, the five differentially expressed miRNAs with the largestdifference multiples were hsa-let-7b-3p, hsa-let-7f-1-3p, hsa-miR-92a-1-5p, hsa-miR-219a-2-3p and hsa-miR-6810-3p, respectively. The results of amplification curve analysis showed that all samples had undergone a good amplification process.The results of melting curve analysis showed that most curves were unimodal. The expression levels of hsa-let-7b-3p and hsamiR-92a-1-5p in the case group were lower than those in the control group, while the expression level of hsa-miR-6810-3pwas higher than that in the control group (P < 0.05) . Conclusion Compared with healthy individuals, patients with idiopathicepilepsy have a total of 98 up-regulated miRNAs and 48 down-regulated miRNAs in their peripheral blood. Among them, the fivedifferentially expressed miRNAs with the largest difference multiples are hsa-let-7b-3p, hsa-let-7f-1-3p, hsa-miR-92a-1-5p,hsa-miR-219a-2-3p, and hsa-miR-6810-3p, respectively. Moreover, only hsa-let-7b-3p, hsa-miR-92a-1-5p and hsa-miR6810-3p might be involved in the occurrence and development of idiopathic epilepsy

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