骨髓间充质干细胞移植联合低剂量雌激素肌肉注射对大鼠脊髓损伤治疗的实验研究

发布时间：2018-08-15 18:19

【摘要】：目前认为,继发性脊髓损伤的分子机制十分复杂,且在病理过程的发生发展中起着重要作用,其中炎症及细胞凋亡是最关键的影响因素,本试验通过观察肌肉注射低剂量雌激素与骨髓间充质干细胞局部移植共同应用对大鼠脊髓损伤后神经运动功能恢复及局部炎症因子变化的影响。骨髓间充质干细胞在某些条件下可定向诱导分化为神经元样细胞、软骨细胞、脂肪细胞、血管内皮细胞等,目前,骨髓间充质干细胞或脂肪干细胞通过各种途径移植治疗急性脊髓损伤的效果己经在较多的基础动物实验和一些临床治疗中获得肯定。雌激素目前已被作为一种抗炎、抗氧化、抑制神经细胞凋亡的生理激素用于实验当中,避免了大剂量糖皮质激素冲击治疗脊髓损伤带来的诸多副作用。研究中我们将100只雄性SD大鼠随机分5组,假手术组仅暴露脊髓,不做损伤处理;模型组、雌激素组、干细胞组、联合组采用改良的Allen's法建立脊髓损伤模型。干细胞组、联合组造模后立即在损伤部位移植骨髓间充质干细胞;雌激素组、联合组造模后第1小时肌肉注射17β-雌二醇。造模后第1,3,5,7天,进行BBB行为学评价;造模后6,12,24,72 h,应用ELISA法测定脊髓匀浆组织白细胞介素1β、肿瘤坏死因子α水平,TUNEL染色观察脊髓组织切片神经细胞凋亡情况。实验结果显示:(1)与假手术组比较,模型组造模后不同时间点的BBB行为学评分下降(P0.05);与模型组比较,干细胞、雌激素组、联合组造模后5,7 d的BBB行为学评分升高(P0.05),且联合组高于干细胞、雌激素组(P0.05);(2)与假手术组比较,模型组造模后6,12,24 h的白细胞介素1β、肿瘤坏死因子α水平升高(P0.05);与模型组比较,干细胞、雌激素组、联合组造模后12,24 h的白细胞介素1β、肿瘤坏死因子α水平降低(P0.05),且联合组低于干细胞、雌激素组(P0.05);(3)造模后72 h,模型组神经细胞凋亡率高于假手术组,干细胞、雌激素组、联合组低于模型组(P0.05),且以联合组最低(P0.05)。雌激素与骨髓间充质干细胞联合应用治疗脊髓损伤具有协同作用,能够更有效抑制神经细胞凋亡,减轻炎症反应,促进大鼠脊髓功能恢复。
[Abstract]:At present, it is believed that the molecular mechanism of secondary spinal cord injury is very complex and plays an important role in the pathogenesis of spinal cord injury. Inflammation and apoptosis are the most important influencing factors. The effects of intramuscular injection of low dose estrogen and local transplantation of bone marrow mesenchymal stem cells on the recovery of neural motor function and the changes of local inflammatory factors after spinal cord injury in rats were observed. Bone marrow mesenchymal stem cells can be induced to differentiate into neuron-like cells, chondrocytes, adipocytes and vascular endothelial cells under certain conditions. The effect of transplantation of bone marrow mesenchymal stem cells or adipose stem cells in the treatment of acute spinal cord injury has been confirmed in many basic animal experiments and some clinical treatments. Estrogen has been used as an anti-inflammatory, anti-oxidative, anti-neuronal apoptosis physiological hormone in the experiment, to avoid a large dose of glucocorticoid shock treatment of spinal cord injury caused by many side effects. In the study, 100 male Sprague-Dawley rats were randomly divided into 5 groups: sham-operated group only exposed spinal cord without injury treatment; Model group, estrogen group, stem cell group, combined group were used to establish spinal cord injury model by modified Allen's method. In the stem cell group, bone marrow mesenchymal stem cells were transplanted at the injured site immediately after modeling in the combined group, and 17 尾 -estradiol was injected intramuscularly in the estrogen group at 1 hour after modeling. The behavior of BBB was evaluated on the 7th day after establishment of the model, and the apoptosis of neurons in spinal cord tissue sections was observed by ELISA assay and Tunel staining. The results showed that: (1) compared with the sham operation group, the BBB behavioral score of the model group decreased at different time points after modeling (P0.05); compared with the model group, stem cells, estrogen group, The BBB behavioral score of the combined group was significantly higher than that of the stem cell group (P0.05), and the level of IL-1 尾 and TNF- 伪 in the estrogen group (P0.05); (2) was significantly higher than that in the sham operation group (P0.05) at 24 h after modeling (P0.05), and was significantly higher than that in the model group (P0.05), and compared with the model group (P0.05), the levels of IL-1 尾 and TNF- 伪 in the model group were significantly higher than those in the sham operation group (P0.05). The levels of IL-1 尾 and TNF- 伪 in stem cells, estrogen group and combination group decreased at 24 h after modeling (P0.05), and were lower in combination group than those in stem cell group. The apoptosis rate of neural cells in model group (P0.05); (3) was higher than that in sham operation group (P 0.05). Estrogen group, the combined group was lower than the model group (P0.05), and the combined group was the lowest (P0.05). Combined use of estrogen and bone marrow mesenchymal stem cells in the treatment of spinal cord injury has a synergistic effect, which can more effectively inhibit neuronal apoptosis, alleviate inflammatory reaction and promote the recovery of spinal cord function in rats.
【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2

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