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外伤性脊髓损伤后移植人乳牙干细胞的神经保护作用涉及抑制早期神经元凋亡。

Neuroprotector effect of stem cells from human exfoliated deciduous teeth transplanted after traumatic spinal cord injury involves inhibition of early neuronal apoptosis.

作者信息

Nicola Fabrício do Couto, Marques Marília Rossato, Odorcyk Felipe, Arcego Danusa Mar, Petenuzzo Letícia, Aristimunha Dirceu, Vizuete Adriana, Sanches Eduardo Farias, Pereira Daniela Pavulack, Maurmann Natasha, Dalmaz Carla, Pranke Patricia, Netto Carlos A

机构信息

Universidade Federal do Rio Grande do Sul, Post Graduate Program in Neuroscience, Porto Alegre, Brazil; Universidade Federal do Rio Grande do Sul, Department of Biochemistry, Porto Alegre, Brazil.

Universidade Federal do Rio Grande do Sul, Post Graduate Program in Neuroscience, Porto Alegre, Brazil; Universidade Federal do Rio Grande do Sul, Department of Biochemistry, Porto Alegre, Brazil.

出版信息

Brain Res. 2017 May 15;1663:95-105. doi: 10.1016/j.brainres.2017.03.015. Epub 2017 Mar 16.

Abstract

UNLABELLED

Stem cells from human exfoliated deciduous teeth (SHED) transplants have been investigated as a possible treatment strategy for spinal cord injuries (SCI) due to their potential for promoting functional recovery. The aim of present study was to investigate the effects of SHED on neuronal death after an experimental model of SCI.

METHODS

Wistar rats were spinalized using NYU impactor®. Animals were randomly distributed into 4 groups: Control (Naive) or Surgical control, Sham (laminectomy with no SCI); SCI (laminectomy followed by SCI, treated with vehicle); SHED (SCI treated with intraspinal transplantation of 3×10 SHED, 1h after SCI). Functional evaluations and morphological analysis were performed to confirm the spinal injury and the benefit of SHED transplantation on behavior, tissue protection and motor neuron survival. Flow cytometry of neurons, astrocytes, macrophages/microglia and T cells of spinal cord tissue were run at six, twenty-four, forty-eight and seventy-two hours after lesion. Six hours after SCI, ELISA and Western Blot were run to assess pro- and anti-apoptotic factors. The SHED group showed a significant functional improvement in comparison to the SCI animals, as from the first week until the end of the experiment. This behavioral protection was associated with less tissue impairment and greater motor neuron preservation. SHED reduced neuronal loss over time, as well as the overexpression of pro-apoptotic factor TNF-α, while maintained basal levels of the anti-apoptotic BCL-XL six hours after lesion. Data here presented show that SHED transplantation one hour after SCI interferes with the balance between pro- and anti-apoptotic factors and reduces early neuronal apoptosis, what contributes to tissue and motor neuron preservation and hind limbs functional recovery.

摘要

未标记

由于人脱落乳牙干细胞(SHED)具有促进功能恢复的潜力,其移植已被研究作为脊髓损伤(SCI)的一种可能治疗策略。本研究的目的是在SCI实验模型后研究SHED对神经元死亡的影响。

方法

使用纽约大学撞击器对Wistar大鼠进行脊髓损伤。动物被随机分为4组:对照组(未处理)或手术对照组、假手术组(行椎板切除术但无SCI);SCI组(行椎板切除术继以SCI,用赋形剂处理);SHED组(SCI后1小时经椎管内移植3×10个SHED进行处理)。进行功能评估和形态学分析以确认脊髓损伤以及SHED移植对行为、组织保护和运动神经元存活的益处。在损伤后6、24、48和72小时对脊髓组织中的神经元、星形胶质细胞、巨噬细胞/小胶质细胞和T细胞进行流式细胞术检测。SCI后6小时,进行ELISA和蛋白质印迹分析以评估促凋亡和抗凋亡因子。与SCI动物相比,SHED组从实验第一周直至实验结束均显示出显著的功能改善。这种行为保护与较少的组织损伤和更多的运动神经元保存相关。随着时间推移,SHED减少了神经元损失以及促凋亡因子TNF-α的过度表达,同时在损伤后6小时维持抗凋亡BCL-XL的基础水平。此处呈现的数据表明,SCI后1小时进行SHED移植可干扰促凋亡和抗凋亡因子之间的平衡并减少早期神经元凋亡,这有助于组织和运动神经元的保存以及后肢功能恢复。

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