Methylprednisolone effects on the migration of endogenous neural stem cells after spinal cord injury

doi:10.3969/j.issn.2095-4344.2016.36.015

Abstract

Abstract:

BACKGROUND: After spinal cord injury, endogenous neural stem cells are activated to proliferate and migrate to repair damaged tissue. As a clinical medicine, methylprednisolone shows a lot of functions, but its effects on endogenous neural stem cells are still unknown.
OBJECTIVE: To explore the effects of methylprednisolone on the proliferation and migration of endogenous neural stem cells after spinal cord injury.
METHODS: Seventy-five Sprague-Dawley rats were used to make animal models of T10 complete paraplegia using Allen’s method, and randomized into methylprednisolone, normal saline and model groups. Rats in these three groups were given intraperitoneal injection of 1 g/L methylprednisolone solution at a dose of 30 mg/kg for 10 minutes and at a dose of 5.4 mg/kg/h for 23 hours, given intraperitoneal injection of normal saline at the same dose and given no treatment, respectively. Neurological and motor functions were assessed by somatosensory evoked potential and Basso Beattie Bresnahan scores at 7, 14, 21, 28 days after spinal cord injury. BrdU and Nestin staining of the injured spinal cord segment was conducted.
RESULTS AND CONCLUSION: A large amount of BrdU- and Nestin-positive cells were visible in all the groups, and the number of these cells reached the peach at 14 days after spinal cord injury. Methylprednisolone was found to inhibit BrdU-, Nestin- or double-positive cells, indicating methylprednisolone can inhibit the proliferation and migration of endogenous neural stem cells. The results of Basso Beattie Bresnahan scores showed no notable improvement in the motor function of the limbs. Methylprednisolone also showed no significant effects on the motor evoked potential latency, but promoted nerve conduction recovery. All these findings indicate that methylprednisolone has some hindering effects on spinal cord repair by inhibiting the proliferation and migration of endogenous neural stem cells after spinal cord injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Spinal Cord Injuries, Neural Stem Cells, Methylprednisolone, Tissue Engineering

CLC Number:

R394.2

Qu Yi-ming, Li Bo, Wang Qun-bo, Shao Gao-hai, Lu Min-peng, Yu Yu, Liu Zuo-zhong, Cao Chun-feng . Methylprednisolone effects on the migration of endogenous neural stem cells after spinal cord injury[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(36): 5419-5425.

Figures/Tables 2

2.1 实验动物数量分析 75只SD大鼠均进入结果分析。 2.2 甲基强的松龙抑制损伤脊髓内BrdU染色的细胞数量 BrdU染色的细胞数量及位置可以用于检测内源神经干细胞的增殖及迁移。在同一倍数的镜下统计BrdU阳性细胞数目(图1)。在正常的大鼠脊髓切片中，大小均匀、圆形或椭圆形的细胞，即表明内源神经干细胞存在。各组BrdU细胞阳性数目见表1。统计分析显示，生理盐水组与模型组差异无显著性意义，甲基强的松龙组各时间点BrdU阳性细胞数均低于生理盐水组，差异有显著性意义(P < 0.05)。 2.3 甲基强的松龙抑制损伤脊髓内Nestin染色的细胞数量 3组各个时间段的Nestin标记的细胞数量与BrdU的变化相同(图2，表2)，均在脊髓损伤后第14天达到最峰，随后降低。Nestin标记的神经干细胞主要集中脊髓损伤的部位。统计分析显示，在各个时间点甲基强的松龙组Nestin表达阳性细胞明显低于生理盐水组，模型组与生理盐水组差异无显著性意义，提示甲基强的松龙能够明显抑制脊髓损伤大鼠的神经干细胞的扩增及迁移。 2.4 甲基强的松龙抑制BrdU+和Nestin+双阳性的细胞数量脊髓损伤后BrdU+和Nestin+双阳性细胞在14 d达到最高，随后降低，该细胞趋向于损伤部位，并且聚集于损伤部位周围(图3)。甲基强的松龙组与生理盐水组相比差异有显著性意义，模型组与生理盐水组差异无显著性意义，见表3。 2.5 甲基强的松龙对于后肢运动无显著性作用利用BBB评分鉴定甲基强的松龙对于脊髓损伤后下肢活动的影响，各组之间比较，差异无显著性意义(P > 0.05)，可见注射甲基强的松龙对于损伤脊髓大鼠后肢运动没有显著作用，见表4。 2.6 脊髓损伤后脊髓运动诱发电位潜伏期和波幅的改变表5显示在各时间点各组大鼠运动诱发电位潜伏期，分析可见各组大鼠运动诱发电位潜伏期比较差异无显著性意义(P > 0.05)，甲基强的松龙对于诱发电位潜伏期没有显著作用。表6显示在各时间点各组大鼠运动诱发电位波幅，可见甲基强的松龙组波幅显著比生理盐水组有所改善，即甲基强的松龙注射可以促进损伤脊髓神经传导的恢复。"

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