Background:The effect of combined repetitive transcranial magnetic stimulation (rTMS) and scalp acupuncture stimulation (SAS) on middle cerebral artery occlusion (MCAO) mice has not yet been reported. The regulation of gene expression after combined stimulation remains unclear. Objective:To analyzegene expression patterns through ribonucleic acid (RNA) sequencing. Methods:Thirty-six 8-weeks-old C57BL/6J male mice weighing 50–60 grams were used for this experiment. The MCAO was induced with 60-min occlusion and subsequent reperfusion of the middle cerebral artery. Experimental mice were randomly assigned to four groups, with nine mice in each group, as follows: control group (no treatment), SAS group (10 minutes SAS), rTMS group (1 Hz rTMS), and combined group (1 Hz rTMS and SAS). Stimulation was performed from the 3rd day to the 7th day after the induction of MCAO. All mice were sacrificed, and brain tissues were taken from the motor area of the MCAO lesion. We analyzed their gene expression profiles using RNA sequencing technology. Results:After stimulation, the grip strength increased in the SAS and rTMS group compared to the control and combined group. The nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) was the key up-regulated protein in the SAS group while src homologus and collagene gene (SHC) and p90 ribosomal protein S6 kinases (p90RSK) were key up-regulated proteins in the rTMS group. However, the C-terminal src kinase-homologous kinase (CHK) was down-regulated whereas p90RSK was up-regulated in the combined group based on the RNA sequencing analysis. Conclusions:Each stimulation method showed different patterns with neurotrophin signaling pathway including NFκB, SHC, p90RSK, and CHK. These can be used in further mechanistic studies about gene expression related to neurorecovery.

中文翻译：

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头针联合经颅磁刺激对大脑中动脉闭塞小鼠基因表达的调控[J].

背景：联合重复经颅磁刺激（rTMS）和头皮针刺激（SAS）对大脑中动脉闭塞（MCAO）小鼠的影响尚未见报道。联合刺激后基因表达的调控仍不清楚。目的：通过核糖核酸（RNA）测序分析基因表达模式。方法：本实验选用36只8周龄C57BL/6J雄性小鼠，体重50～60克。MCAO 是通过 60 分钟的闭塞和随后的大脑中动脉再灌注诱导的。实验小鼠随机分为四组，每组9只，分别为：对照组（未治疗）、SAS组（10分钟SAS）、rTMS组（1Hz rTMS）和联合组（1Hz rTMS和SAS）。诱导MCAO后第3天至第7天进行刺激。处死所有小鼠，并从 MCAO 病变的运动区取出脑组织。我们使用 RNA 测序技术分析了他们的基因表达谱。结果：刺激后，SAS和rTMS组的握力较对照组和联合组增加。活化 B 细胞的核因子 kappa 轻链增强子 (NFκB) 是 SAS 组中关键的上调蛋白，而 src 同源和胶原基因 (SHC) 和 p90 核糖体蛋白 S6 激酶 (p90RSK) 是关键上调蛋白rTMS 组中的调节蛋白。然而，基于 RNA 测序分析，C 端 src 激酶同源激酶 (CHK) 被下调而 p90RSK 在组合组中被上调。结论：每种刺激方法都显示出不同的神经营养因子信号通路模式，包括 NFκB、SHC、p90RSK 和 CHK。这些可用于与神经恢复相关的基因表达的进一步机制研究。