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羊水间充质干细胞分泌的神经营养因子促进大鼠坐骨神经损伤后再生

Post-injury regeneration in rat sciatic nerve facilitated by neurotrophic factors secreted by amniotic fluid mesenchymal stem cells.

作者信息

Pan Hung-Chuan, Cheng Fu-Chou, Chen Chun-Jung, Lai Shu-Zhen, Lee Chi-Wen, Yang Dar-Yu, Chang Ming-Hong, Ho Shu-Peng

机构信息

Department of Neurosurgery, Taichung Veterans General Hospital, Taichung, Taiwan.

出版信息

J Clin Neurosci. 2007 Nov;14(11):1089-98. doi: 10.1016/j.jocn.2006.08.008.

Abstract

Amniotic fluid mesenchymal stem cells have the ability to secrete neurotrophic factors that are able to promote neuron survival in vitro. The purpose of this study was to evaluate the effects of neurotrophic factors secreted by rat amniotic fluid mesenchymal stem cells on regeneration of sciatic nerve after crush injury. Fifty Sprague-Dawley rats weighing 250-300 g were used. The left sciatic nerve was crushed with a vessel clamp. Rat amniotic fluid mesenchymal stem cells embedded in fibrin glue were delivered to the injured nerve. Enzyme-linked immunosorbent assay (ELISA) and immunocytochemistry were used to detect neurotrophic factors secreted by the amniotic fluid mesenchymal stem cells. Nerve regeneration was assessed by motor function, electrophysiology, histology, and immunocytochemistry studies. Positive CD29/44, and negative CD11b/45, as well as high levels of expression of brain-derived neurotrophic factor, glia cell line-derived neurotrophic factor, ciliary neurotrophic factor (CNTF), nerve growth factor, and neurotrophin-3 (NT-3) were demonstrated in amniotic fluid mesenchymal stem cells. Motor function recovery, the compound muscle action potential, and nerve conduction latency showed significant improvement in rats treated with amniotic fluid mesenchymal stem cells. ELISA measurement in retrieved nerves displayed statistically significant elevation of CNTF and NT-3. The immunocytochemical studies demonstrated positive staining for NT-3 and CNTF in transplanted cells. The histology and immunocytochemistry studies revealed less fibrosis and a high level of expression of S-100 and glial fibrillary acid protein at the crush site. Rat amniotic fluid mesenchymal stem cells may facilitate regeneration in the sciatic nerve after crush injury. The increased nerve regeneration found in this study may be due to the neurotrophic factors secreted by amniotic fluid mesenchymal stem cells.

摘要

羊水间充质干细胞具有分泌神经营养因子的能力,这些神经营养因子能够在体外促进神经元存活。本研究的目的是评估大鼠羊水间充质干细胞分泌的神经营养因子对坐骨神经挤压伤后再生的影响。使用了50只体重250 - 300克的Sprague-Dawley大鼠。用血管夹夹伤左侧坐骨神经。将包埋于纤维蛋白胶中的大鼠羊水间充质干细胞移植到受损神经处。采用酶联免疫吸附测定(ELISA)和免疫细胞化学方法检测羊水间充质干细胞分泌的神经营养因子。通过运动功能、电生理学、组织学和免疫细胞化学研究评估神经再生情况。在羊水间充质干细胞中证实有CD29/44阳性、CD11b/45阴性,以及脑源性神经营养因子、胶质细胞源性神经营养因子、睫状神经营养因子(CNTF)、神经生长因子和神经营养素-3(NT-3)的高表达。在接受羊水间充质干细胞治疗的大鼠中,运动功能恢复、复合肌肉动作电位和神经传导潜伏期均有显著改善。对取出神经的ELISA检测显示CNTF和NT-3有统计学意义的升高。免疫细胞化学研究显示移植细胞中NT-3和CNTF呈阳性染色。组织学和免疫细胞化学研究显示挤压部位纤维化较少,S-100和胶质纤维酸性蛋白表达水平较高。大鼠羊水间充质干细胞可能促进坐骨神经挤压伤后的再生。本研究中发现的神经再生增加可能归因于羊水间充质干细胞分泌的神经营养因子。

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