Department of Orthopedics, Renmin Hospital of Wuhan University, Wuchang District, Wuhan, Hubei Province 430060, China.
Department of Chinese Traditional Medicine, Tongren Hospital of Wuhan University(Wuhan Third Hospital), Wuchang District, Wuhan, Hubei Province 430060, China.
Gene. 2021 May 20;781:145528. doi: 10.1016/j.gene.2021.145528. Epub 2021 Feb 22.
Spinal cord injury (SCI) leads to severe motor and sensory dysfunctions. Neural stem cells (NSCs) transplantation therapy plays a positive role in functional recovery after SCI, but the effectiveness of this therapy is limited by inadequate differentiation ability of transplanted NSCs. Mammalian achaete-scute homologue-1 (Mash-1) has been reported to improve differentiation of NSCs. Thus, this study modified NSCs with Mash-1 to repair SCI.
NSCs isolated from rat embryo hippocampus were cultured and identified in vitro and further transfected with the lentiviral vectors (Lv-Mash-1). After establishing a SCI rat model, the rats were transplanted with Mash-1 modified NSCs, the histopathological changes of rat spinal cord were detected by hematoxylin-eosin (HE) staining, and the locomotor activity of rats was evaluated with the Basso, Beattie and Bresnahan (BBB) scale. The NSCs cultured in vitro or extracted from SCI rat spinal cord were identified by immunofluorescence (IF). Mash-1, β3-Tubulin, and NeuN expressions in those cells were determined by Western blotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR).
NSCs isolated from rat embryo hippocampus were Nestin- and NeuN-positive. NSC transplantation modified by Mash-1 increased BBB score of SCI rats and promoted recovery in lesion site of SCI rats. Mash-1 overexpression also promoted β3-Tubulin and NeuN expressions in NSCs cultured in vitro or extracted from spinal cord of SCI rats.
Mash-1 overexpression promoted NSC differentiation into neurons, and further improved locomotor functional recovery of SCI rats.
脊髓损伤（SCI）导致严重的运动和感觉功能障碍。神经干细胞（NSCs）移植疗法在 SCI 后的功能恢复中发挥积极作用，但该疗法的有效性受到移植 NSCs 分化能力不足的限制。哺乳动物achaete-scute 同源物-1（Mash-1）已被报道可改善 NSCs 的分化能力。因此，本研究通过修饰 NSCs 中的 Mash-1 来修复 SCI。
体外培养和鉴定大鼠胚胎海马来源的 NSCs，并进一步用慢病毒载体（Lv-Mash-1）转染。建立 SCI 大鼠模型后，将 Mash-1 修饰的 NSCs 移植入大鼠，通过苏木精-伊红（HE）染色检测大鼠脊髓的组织病理学变化，Basso、Beattie 和 Bresnahan（BBB）评分评估大鼠的运动活动。通过免疫荧光（IF）鉴定体外培养的 NSCs 或从 SCI 大鼠脊髓中提取的 NSCs。通过 Western blot 和逆转录-定量聚合酶链反应（RT-qPCR）测定这些细胞中 Mash-1、β3-Tubulin 和 NeuN 的表达。
从大鼠胚胎海马中分离的 NSCs 呈 Nestin 和 NeuN 阳性。Mash-1 修饰的 NSC 移植增加了 SCI 大鼠的 BBB 评分，并促进了 SCI 大鼠损伤部位的恢复。Mash-1 的过表达也促进了体外培养的 NSCs 或从 SCI 大鼠脊髓中提取的 NSCs 中β3-Tubulin 和 NeuN 的表达。
Mash-1 的过表达促进了 NSCs 向神经元的分化，并进一步改善了 SCI 大鼠的运动功能恢复。
