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Overexpression of miR-124 in astrocyte improves neurological deficits in rat with ischemic stroke via DLL4 modulation

纽恩 转染 星形胶质细胞 重编程 生物 小胶质细胞 基因敲除 胶质瘢痕 双皮质醇 病毒载体 神经元 细胞生物学 分子生物学 细胞 神经科学 免疫学 免疫组织化学 细胞培养 中枢神经系统 炎症 生物化学 基因 齿状回 遗传学 重组DNA
作者
Xiaoqin Guo,Pingli Jiang,Meihua Pan,Yanyi Ding,Yanting Lin,Tao Jiang,Rui Li,Wen‐ju Wang,Yaling Dai,Sinuo Wang,Yajun Cao,Huawei Lin,Minguang Yang,Weilin Liu,Jing Tao
出处
期刊:Experimental Neurology [Elsevier]
卷期号:370: 114571-114571 被引量:2
标识
DOI:10.1016/j.expneurol.2023.114571
摘要

Astrocytes have been demonstrated to undergo conversion into functional neurons, presenting a promising approach for stroke treatment. However, the development of small molecules capable of effectively inducing this cellular reprogramming remains a critical challenge. Initially, we introduced a glial cell marker gene, GFaABC1D, as the promoter within an adeno-associated virus vector overexpressing miR-124 into the motor cortex of an ischemia-reperfusion model in rats. Additionally, we administered NeuroD1 as a positive control. Lentiviral vectors overexpressing miR-124 were constructed and transfected into primary rat astrocytes. We assessed the cellular distribution of GFAP, DCX, and NeuN on days 7, 14, and 28, respectively. In rats with ischemic stroke, miR-124-transduced glial cells exhibited positive staining for the immature neuron marker doublecortin (DCX) and the mature neuron marker NeuN after 4 weeks. In contrast, NeuroD1-overexpressing model rats only expressed NeuN, and the positive percentage was higher in co-transfection with miR-124 and NeuroD1. Overexpression of miR-124 effectively ameliorated neurological deficits and motor functional impairment in the model rats. In primary rat astrocytes transduced with miR-124, DCX was not observed after 7 days of transfection, but it appeared at 14 days, with the percentage further increasing to 44.6% at 28 days. Simultaneously, 15.1% of miR-124-transduced cells exhibited NeuN positivity, which was not detected at 7 and 14 days. In vitro, double fluorescence assays revealed that miR-124 targeted Dll4, and in vivo experiments confirmed that miR-124 inhibited the expression of Notch1 and DLL4. The overexpression of miR-124 in astrocytes demonstrates significant potential for improving neurological deficits following ischemic stroke by inhibiting DLL4 expression, and it may facilitate astrocyte-to-neuronal transformation.
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