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Effects of Edaravone on Functional Recovery of a Rat Model with Spinal Cord Injury Through Induced Differentiation of Bone Marrow Mesenchymal Stem Cells into Neuron-Like Cells
Cellular Reprogramming ( IF 1.6 ) Pub Date : 2021-02-02 , DOI: 10.1089/cell.2020.0055
Yumei Li 1 , Laibing Liu 2 , Zijiang Yu 1 , Yan Yu 1 , Baofei Sun 1 , Chaolun Xiao 1 , Shipeng Luo 1 , Lin Li 1
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Edaravone can induce differentiation of bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells and replace lost cells by transplanting neuron-like cells to repair spinal cord injury (SCI). In this study, BMSCs were derived from the bone marrow of male Wistar rats (4 weeks old) through density gradient centrifugation (1.073 g/mL), and the cell purity of BMSCs was up to 95%. The combined injection of basic fibroblast growth factor and edaravone was conducted to differentiate BMSCs into neuron-like cells. In this study, 120 male Wistar rats were used to establish the model of semitransverse SCI; on the seventh day, neuron-like cells were labeled by BrdU and then injected into the epicenter of the injury of rats. On the 14th day after cell transplantation, the biotin dextran amine (BDA) fluorescent agent was used to track the repair of nerve damage. At 7, 14, 21, and 30 days after SCI, the Basso, Beattie, and Bresnahan (BBB) locomotor scale method was used to measure the functional recovery of hind limbs in rats. Additionally, hematoxylin and eosin (H&E) staining, Nissl staining, immunohistochemistry, transmission electron microscopy (TEM), Western blotting, and Real-time quantitative reverse transcripion PCR (qRT-PCR) were used to observe the regeneration of nerve cells. In the edaravone+BMSC group, behavioral analysis of locomotor function showed that functional recovery was significantly enhanced after transplantation of the cells, BrdU-positive cells could be observed scattered in the injured area and extended to both the head and tail, and the BDA tracer shows that the edaravone+BMSC group emits more fluorescent signals. Additionally, H&E staining, Nissl staining, and immunohistochemistry revealed that the space of spinal cord tissue was attenuated and the neurons were increased. Western blotting and qRT-PCR showed that the expression levels of neuron-specific enolase (NSE), Nestin, and neurofilament 200 (NF) were increased, while the expression of glial fibrillary acidic protein (GFAP) was decreased. TEM showed that cytoplasmic edema was reduced, mitochondrial vacuoles were attenuated, and nuclear chromatin concentration was declined after transplantation of neuron-like cells. Moreover, with the extension of time of edaravone+BMSC transplantation, the structures of mitochondria and endoplasmic reticulum tended to be normal. In summary, the induced differentiation of BMSC transplantation can significantly promote the functional repair of SCI.

中文翻译:

依达拉奉通过诱导骨髓间充质干细胞向神经元样细胞分化对脊髓损伤大鼠模型功能恢复的影响

依达拉奉可诱导骨髓间充质干细胞(BMSCs)分化为神经元样细胞,并通过移植神经元样细胞替代丢失的细胞来修复脊髓损伤(SCI)。在本研究中,BMSCs 从雄性 Wistar 大鼠(4 周龄)的骨髓中通过密度梯度离心(1.073 g/mL),骨髓间充质干细胞的细胞纯度高达 95%。碱性成纤维细胞生长因子和依达拉奉联合注射使BMSCs分化为神经元样细胞。本研究采用120只雄性Wistar大鼠建立半横断脊髓损伤模型;第7天,神经元样细胞用BrdU标记,然后注射到大鼠损伤的震中。细胞移植后第14天,使用生物素葡聚糖胺(BDA)荧光剂追踪神经损伤的修复情况。在 SCI 后 7、14、21 和 30 天,采用 Basso、Beattie 和 Bresnahan (BBB) 运动量表法测量大鼠后肢的功能恢复情况。此外,苏木精和伊红 (H&E) 染色、尼氏染色、免疫组织化学、透射电子显微镜 (TEM)、蛋白质印迹、实时定量逆转录PCR(qRT-PCR)用于观察神经细胞的再生。依达拉奉+BMSC组运动功能行为学分析显示细胞移植后功能恢复明显增强,可观察到BrdU阳性细胞散布于损伤部位并延伸至头尾,BDA示踪剂显示依达拉奉+BMSC组发出更多的荧光信号。此外,H&E染色、Nissl染色和免疫组化显示脊髓组织间隙变薄,神经元增多。Western印迹和qRT-PCR显示神经元特异性烯醇化酶(NSE)、巢蛋白和神经丝200(NF)的表达水平升高,而胶质纤维酸性蛋白(GFAP)的表达降低。TEM显示神经元样细胞移植后细胞质水肿减轻,线粒体空泡减弱,核染色质浓度下降。此外,随着依达拉奉+骨髓间充质干细胞移植时间的延长,线粒体和内质网结构趋于正常。综上所述,骨髓间充质干细胞移植诱导分化可显着促进脊髓损伤的功能修复。线粒体和内质网结构趋于正常。综上所述,骨髓间充质干细胞移植诱导分化可显着促进脊髓损伤的功能修复。线粒体和内质网结构趋于正常。综上所述,骨髓间充质干细胞移植诱导分化可显着促进脊髓损伤的功能修复。
更新日期:2021-02-03
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