Qiao Chenye, Ran Yuanyuan, Li Ning, Wang Congxiao, Li Jinglu, Xi Xiaoming, Li Zihan, Ye Lin, Su Wei, Liu Zongjian, Qie Shuyan
Beijing Rehabilitation Hospital, Capital Medical University, Beijing 100144, China.
Beijing International Science and Technology Cooperation Base for Antiviral Drugs, College of Chemistry and Life Science, Beijing University of Technology, Beijing 100124, China.
Exp Neurol. 2025 Jul;389:115255. doi: 10.1016/j.expneurol.2025.115255. Epub 2025 Apr 10.
Neuroplasticity is crucial for functional recovery after stroke, with modulation of microglial polarization enhancing this process. Intermittent theta burst stimulation (iTBS), as a neuromodulation technique, can simultaneously generate electric and magnetic fields to act on the central nervous system. Neurons can induce electrochemical signal transduction as excitable cells. Meanwhile, iTBS can regulate microglial inflammatory polarization post-stroke. However, how iTBS exerts its effect on microglia remains unclear. The magnetoreceptive protein Cryptochrome (Cry) can respond to the magnetic effect and is known to regulate macrophage-mediated inflammatory responses. However, whether iTBS modulates microglial polarization through Cry1 is unknown.
To explore the magnetic effects of iTBS on microglial polarization through Cry1, thereby enhancing neuroplasticity and stroke recovery, and also elucidate the role of the Cry1-NF-κB pathway in iTBS-mediated regulation of microglial polarization.
A mouse model was established using photothrombosis (PT), followed by 7-day iTBS intervention. BV2 cells and primary neurons were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) respectively, followed by once-daily iTBS treatment for two days. Brain damage and functional recovery were assessed using Map-2 staining and behavioral tests. RT-PCR, western blot, immunofluorescence and transwell co-culture experiments were employed to evaluate the effects of iTBS on microglial polarization and neuroplasticity. Cry1 knockdown via siRNA transfection was used to explore the Cry1-NF-κB signaling pathway.
iTBS ameliorated neuronal damage induced by ischemic injury, reduced pro-inflammatory microglial activation, and promoted anti-inflammatory polarization. Cry1 expression was upregulated in BV2 cells in response to iTBS, while Cry1 knockdown increased CD16 expression, decreased CD206 expression and further alleviate the inhibition of NF-κB activation. In primary neurons, anti-inflammatory microglia induced by iTBS could enhance neuroplasticity.
This study demonstrates that post-stroke iTBS promotes neuroplasticity and functional recovery by regulating microglial polarization via the Cry1-NF-κB pathway.
神经可塑性对中风后的功能恢复至关重要，小胶质细胞极化的调节可增强这一过程。间歇性theta爆发刺激（iTBS）作为一种神经调节技术，可同时产生电场和磁场作用于中枢神经系统。神经元作为可兴奋细胞可诱导电化学信号转导。同时，iTBS可调节中风后小胶质细胞的炎症极化。然而，iTBS如何对小胶质细胞发挥作用仍不清楚。磁感受蛋白隐花色素（Cry）可对磁效应做出反应，且已知其可调节巨噬细胞介导的炎症反应。然而，iTBS是否通过Cry1调节小胶质细胞极化尚不清楚。
探讨iTBS通过Cry1对小胶质细胞极化的磁效应，从而增强神经可塑性和促进中风恢复，并阐明Cry1-NF-κB通路在iTBS介导的小胶质细胞极化调节中的作用。
采用光血栓形成（PT）建立小鼠模型，随后进行为期7天的iTBS干预。BV2细胞和原代神经元分别进行氧糖剥夺/再灌注（OGD/R），然后每天进行一次iTBS处理，持续两天。使用Map-2染色和行为测试评估脑损伤和功能恢复情况。采用RT-PCR、蛋白质免疫印迹、免疫荧光和Transwell共培养实验评估iTBS对小胶质细胞极化和神经可塑性的影响。通过siRNA转染敲低Cry1以探索Cry1-NF-κB信号通路。
iTBS改善了缺血性损伤诱导的神经元损伤，减少了促炎性小胶质细胞的激活，并促进了抗炎极化。iTBS刺激后BV2细胞中Cry1表达上调，而Cry1敲低增加了CD16表达，降低了CD206表达，并进一步减轻了对NF-κB激活的抑制。在原代神经元中，iTBS诱导的抗炎性小胶质细胞可增强神经可塑性。
本研究表明，中风后iTBS通过Cry1-NF-κB通路调节小胶质细胞极化，从而促进神经可塑性和功能恢复。
