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脂肪来源干细胞联合丝素蛋白/壳聚糖支架过表达脑源性神经营养因子和神经营养素-3对脊髓损伤的保护作用

Protective effect of brain-derived neurotrophic factor and neurotrophin-3 overexpression by adipose-derived stem cells combined with silk fibroin/chitosan scaffold in spinal cord injury.

作者信息

Ji Wen-Chen, Li Meng, Jiang Wan-Ting, Ma Xing, Li Jia

机构信息

Department of Orthopaedics, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.

Department of Ultrasound Diagnosis, The Fourth Hospital of Xi'an, Xi'an, China.

出版信息

Neurol Res. 2020 May;42(5):361-371. doi: 10.1080/01616412.2020.1735819. Epub 2020 Mar 9.

Abstract

: Spinal cord injury (SCI) is a most debilitating traumatic injury, and cytotherapy is a promising alternative treatment strategy. Here we investigated the effect and mechanism of adipose-derived stem/stromal cells (ASCs) with overexpressing brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) (BDNF-NT3) in combination with silk fibroin/chitosan scaffold (SFCS) in SCI.: Female Sprague-Dawley rats were used as an SCI model. SFCS,SFCS and ASCs, or ASCs overexpressing NT3, BDNF, and BDNF-NT3 were implanted into SCI rats. Basso, Beattie, and Bresnahan score, pathological changes, and spinal cord tissue and nerve fiber morphology were observed and assayed. GAP-43, GFAP, and caspase-3 expression was determined using immunohistochemistry and western blotting.: Smoother spinal cords, less scar tissue, and lower inflammatory activity were found in the SFCS, SFCS and ASCs, ASCs with NT3, BDNF, and BDNF-NT3 overexpression treatment than in the untreated SCI rat groups. Increasing formation of nerve fibers was observed in the above groups in order. GAP-43 expression significantly increased, while GFAP and caspase-3 expression significantly decreased. These results indicated obvious alleviation in pathological changes and BDNF-NT3 overexpression in ASCs combined with SFCS treatment in SCI rats.: Thus, BDNF-NT3 overexpression from ASCs with SFCS had synergistic neuroprotective effects on SCI and may be a treatment option for SCI.

摘要

脊髓损伤(SCI)是一种极具致残性的创伤性损伤,而细胞疗法是一种有前景的替代治疗策略。在此,我们研究了过表达脑源性神经营养因子(BDNF)和神经营养因子-3(NT3)(BDNF-NT3)的脂肪来源干细胞/基质细胞(ASCs)与丝素蛋白/壳聚糖支架(SFCS)联合应用于脊髓损伤的效果及机制。

选用雌性Sprague-Dawley大鼠作为脊髓损伤模型。将SFCS、SFCS与ASCs、或过表达NT3、BDNF和BDNF-NT3的ASCs植入脊髓损伤大鼠体内。观察并测定Basso、Beattie和Bresnahan评分、病理变化以及脊髓组织和神经纤维形态。采用免疫组织化学和蛋白质印迹法测定生长相关蛋白43(GAP-43)、胶质纤维酸性蛋白(GFAP)和半胱天冬酶-3(caspase-3)的表达。

与未治疗的脊髓损伤大鼠组相比,SFCS组、SFCS与ASCs联合组、过表达NT3、BDNF和BDNF-NT3的ASCs治疗组的脊髓更光滑,瘢痕组织更少,炎症活性更低。上述各组依次观察到神经纤维形成增加。GAP-43表达显著增加,而GFAP和caspase-3表达显著降低。这些结果表明,ASCs联合SFCS治疗可明显减轻脊髓损伤大鼠的病理变化并使BDNF-NT3过表达。

因此,ASCs联合SFCS过表达BDNF-NT3对脊髓损伤具有协同神经保护作用,可能是脊髓损伤的一种治疗选择。

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