CIMT通过调控c-Jun/miR-182-5p/Nogo-A轴促进偏瘫小鼠神经重塑的机制研究

Constraint-induced movement therapy (CIMT) could improve manual ability in children with hemiplegic cerebral palsy (HCP), which may be related to the mechanism of CIMT-induced neural remodeling. As previous studies and our early researches showed that CIMT could inhibit the expression of Nogo-A to promote neural remodeling. We had further found that CIMT could increase the expression of miR-182-5p and correspondingly inhibit Nogo-A. However, The upstream mechanism of miR-182-5p regulated by CIMT is unclear. According to the bioinformatics websites, transcription factor c-Jun may bind to miR-182-5p, and it was confirmed that c-Jun involved in neural remodeling. It was found that physical stimulation could activate the expression of transcription factor. Based on the above researches, we hypothesized that, CIMT could activate transcription factor c-Jun to activate the transcription of miR-182-5p and inhibit the Nogo-A, which finally improve the neural remodeling in HCP mice. The project is intended to conduct cellular and animal experiments using CHIP, siRNA, lentiviral transfection and neural tracing techniques to reveal the mechanism of CIMT promoting neuroremodeling by regulating c-Jun/miR-182-5p/Nogo-A in hemiplegia mice.This research may offer powerful evidence for the promotion of CIMT and improve the precise therapy for patients of HCP.

强制性运动疗法（CIMT）可促进偏瘫型脑瘫（HCP）患儿手功能康复，其机制与CIMT诱导神经重塑有关。文献及我们前期研究证实，CIMT可抑制Nogo-A表达促进神经重塑。我们进一步发现CIMT可上调miR-182-5p表达并抑制Nogo-A。然而，CIMT调控miR-182-5p的上游机制不清。生物信息学预测转录因子c-Jun可结合miR-182-5p，而c-Jun参与中枢及外周神经重塑的作用已证实。研究提示物理刺激可促进转录因子表达。为此我们推测CIMT可激活转录因子c-Jun，启动miR-182-5p转录，从而抑制Nogo-A，最终参与HCP小鼠神经重塑。本项目开展细胞及动物实验，借助CHIP，siRNA，慢病毒转染和神经示踪等技术，阐明CIMT调控c-Jun/miR-182-5p/Nogo-A轴促进HCP小鼠神经重塑的作用机制，为CIMT的临床推广及HCP的精准治疗提供证据支持。

强制性运动疗法（CIMT）可促进偏瘫型脑瘫（HCP）患儿手功能康复，其机制与CIMT诱导神经重塑有关。文献及我们前期研究证实，CIMT可抑制Nogo-A表达促进神经重塑。我们进一步发现CIMT可上调miR-182-5p表达并抑制Nogo-A。然而，CIMT调控miR-182-5p的上游机制不清。为此，本研究在前期研究的基础上，旨在明确CIMT通过调控c-Jun/miR-182-5p/Nogo-A轴促进HCP小鼠神经重塑的分子机制。本研究通过开展Neuro-2a细胞实验和构建HCP小鼠模型，借助siRNA，慢病毒转染、行为学检测、免疫荧光染色、高尔基染色、透射电镜及蛋白免疫印迹等技术，检测各组小鼠运动功能变化、神经元和神经纤维密度、树突/轴突面积、髓鞘完整性和大脑运动皮层中c-Jun，miR-182-5p，Nogo-A的表达情况，以及c-Jun启动miR-182-5p的转录情况。本研究细胞实验研究证实，转录因子c-Jun可通过结合miR-182-5p以激活其转录；动物研究进一步研究发现，CIMT可改善HCP小鼠的前肢抓握能力和平衡协调能力，并减轻HCP小鼠大脑树突棘和髓鞘损伤，促进神经元再生；CIMT还可上调小鼠脑内c-Jun表达，启动miR-182-5p转录，进而抑制Nogo-A表达以促进HCP小鼠大脑神经重塑，最终改善HCP小鼠肢体运动功能。本项目的研究结果与预期实验结果相符，不仅为CIMT在HCP中的应用推广提供了坚实的理论依据，也为HCP的精准治疗提供了新思路。