陈建敏,李菁,黄华垚,等.基于微小核糖核酸-信使核糖核酸网络探讨重复经颅磁刺激治疗缺血性脑卒中的分子机制[J].中华物理医学与康复杂志,2024,46(7):593-600
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| 基于微小核糖核酸-信使核糖核酸网络探讨重复经颅磁刺激治疗缺血性脑卒中的分子机制 |
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| DOI：10.3760/cma.j.issn.0254-1424.2024.07.003 |
| 中文关键词: 缺血性脑卒中 生物信息学 重复经颅磁刺激 基因表达综合数据库数据库 微小核糖核酸-信使核糖核酸调控网络 |
| 英文关键词: Ischemia Stroke Bioinformatics Transcranial magnetic stimulation GEO database miRNA-mRNA regulation |
| 基金项目:福建省科技创新联合资金引领项目（2020Y9110；2023Y9133）；福建医科大学启航基金（2019QH1026） |
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| 中文摘要: |
| 目的 基于微小核糖核酸（miRNA）-信使核糖核酸（mRNA）调控网络探讨重复经颅磁刺激（rTMS）促进缺血性脑卒中（IS）神经修复的潜在分子机制。 方法 从基因表达综合数据库（GEO）下载rTMS干预7 d后的IS大鼠皮质miRNA表达谱。使用R软件分析差异miRNAs，通过在线数据库预测其对应的下游mRNAs，构建miRNA-mRNA调控网络，筛选关键miRNAs。对靶基因进行功能富集分析，构建靶基因所编码的蛋白质相互作用网络，识别网络中的核心mRNAs。选取无特定病原体动物（SPF）级雄性Sprague-Dawley大鼠24只，按随机数字表法随机分为假手术组、模型组和磁刺激组，每组8只大鼠。模型组和磁刺激组制备IS大鼠模型，仅磁刺激组接受连续7 d的rTMS干预，其余2组不进行干预。采用改良神经功能评分（mNSS）于造模前和造模成功1 d、3 d、7 d后评价3组大鼠的神经功能缺损程度；于造模成功8 d后深度麻醉大鼠，取脑组织行苏木精-伊红和尼氏染色观察组织缺失情况和神经元形态，利用实时荧光定量聚合酶链反应验证差异基因在3组大鼠缺血侧皮质中的表达趋势。 结果 共筛选出167个差异miRNAs和预测到25个mRNAs。富集分析表明，这些基因与细胞迁移的正向调节、神经营养因子受体信号通路的正向调节等生物学过程有关，涉及腺苷酸激活蛋白激酶、神经营养因子通路、大鼠肉瘤等信号通路。通过构建miRNA-mRNA调控网络，识别出miR-206-3p、miR-378a-3p、miR-107-3p、miR-92a-3p和miR-29b-3p共5个关键miRNAs；通过蛋白互作网络分析筛选出4个核心mRNAs，分别为整合素亚基β1、水通道蛋白4、脑源性神经生长因子（BDNF）、溶质载体家族2成员4。造模成功7 d后，磁刺激组的mNSS评分显著低于模型组同时间点（P＜0.05）。与模型组相比，磁刺激大鼠右侧皮质的miR-206-3p表达明显降低（P＜0.05），BDNF表达则显著增高（P＜0.05）。 结论 miR-206-3p-BDNF调控网络和相关作用途径在rTMS促进IS大鼠神经修复过程中发挥重要的作用。 |
| 英文摘要: |
| Objective To explore the molecular mechanism through which repetitive transcranial magnetic stimulation (rTMS) promotes nerve regeneration after ischemic stroke. Methods The miRNA expression profile in rats after rTMS was downloaded from the GEO database. Differentially-expressed miRNAs were identified using R software, and their corresponding mRNAs were predicted using the online database in order to construct miRNA-mRNA regulatory networks and identify the key miRNAs. The protein interaction network encoded by each gene was constructed by functional enrichment analysis, and the core mRNAs in the network were identified. Twenty-four male Sprague-Dawley rats of a specific pathogen-free grade were randomly divided into a sham operation group, a model group and a magnetic stimulation group, each of 8, and a stroke model was induced in the model and magnetic stimulation groups. The magnetic stimulation group then received rTMS for 7 consecutive days, while the other 2 groups did not. Modified neural function scores (mNSSs) were used to quantify neural function deficits before the experiment and 1, 3 and 7 days after the modeling. Eight days after the modeling, brain tissues were sampled for hematoxylin-eosin and Nishin staining to observe tissue loss and neuron morphology. Real-time fluorescence quantitative polymerase chain reactions were employed to verify the differential gene expression in the ischemic cortex. Results A total of 167 different miRNAs were screened, and 25 mRNAs were predicted. Enrichment analysis showed that those genes are related to the positive regulation of cell migration and the positive regulation of neurotrophic factor receptor signaling pathways, including adenylate-activated protein kinase, the neurotrophic factor pathway, and rat sarcoma signaling pathways. The miRNA-mRNA regulatory network highlighted miR-206-3p, miR-378a-3p, miR-107-3p, miR-92a-3p and miR-29b-3p as key miRNAs. Integrin subunit β1, aquaporin 4, brain-derived nerve growth factor (BDNF), and member 4 of solutes vector family 2 were identified as key mRNAs by the protein interaction network analysis. Seven days after the modeling, the average mNSS score of the magnetic stimulation group was significantly lower than the model group′s average. Compared with the model group, the expression of miR-206-3p in the right cortex of the magnetic stimulation group had decreased significantly, while BDNF expression had increased significantly. Conclusions miR-206-3p-BDNF pathways play an important role in neural repair promoted by rTMS. |
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