SHH信号通路成分在成年大鼠脊髓损伤后的表达研究
发布时间：2019-06-22 12:47
【摘要】：目的：在大鼠脊髓组织受到急性损伤后,观察脊髓内源性神经前体细胞的增殖与分化规律,观察与神经前体细胞生长发育相关的Sonic Hedgehog (SHH)信号通路成分SHH和Gli-1分子的表达变化,探讨成年大鼠脊髓损伤后内源性神经前体细胞分化调控的分子机制过程。方法：构建大鼠脊髓急性损伤模型,运用随机数表法将健康成年大鼠分为正常组,假手术组与脊髓损伤各时间组,用5-溴-2-脱氧尿嘧啶核苷(BrdU)示踪剂、胶质原纤维酸性蛋白(glial fibrillary acidic protein、GFAP)、髓鞘碱性蛋白(myelin basic protein,MBP)分别标记神经前体细胞、星形胶质细胞与少突胶质细胞,通过免疫荧光技术观察不同组BrdU标记阳性细胞数和BrdU/GFAP、 BrdU/MBP双标阳性细胞数目的变化趋势,探讨脊髓损伤后神经前体细胞的增殖分化规律。此外,应用实时荧光定量PCR技术和Western blotting技术检测大鼠各组SHH信号通路成分SHH和Gli-1信号分子的变化趋势,探讨SHH信号通路在大鼠脊髓损伤后的调控作用。结果：对于神经前体细胞的研究发现,正常组及假手术组大鼠脊髓均可观察到少量BrdU阳性细胞以及BrdU/GFAP、BrdU/MBP双标阳性细胞,而脊髓损伤后BrdU阳性细胞与双标阳性细胞的表达呈现出显著增高的趋势(P0.05)。此外,对于SHH信号通路成分的研究发现,正常大鼠脊髓低度表达SHH和Gli-1信号分子,脊髓损伤后SHH和Gli-1的表达量持续升高,并在损伤后第7天达到峰值,脊髓损伤后第21天仍然处于高水平表达(P0.05)。结论：大鼠脊髓损伤神经前体细胞增殖明显增多并向星形胶质细胞与少突胶质细胞大量分化。在健康正常大鼠可观察到SHH信号转导通路成分的表达,脊髓损伤后,SHH和Gli-1蛋白的表达明显增多,其变化规律与神经前体细胞的增殖变化规律基本一致,提示SHH信号通路可能参与脊髓损伤后神经细胞修复的调控作用。
[Abstract]:Aim: to observe the proliferation and differentiation of intrinsic neural progenitor cells in spinal cord after acute spinal cord injury, to observe the expression of SHH and Gli-1, which are related to the growth and development of neural progenitor cells, and to explore the molecular mechanism of differentiation regulation of endogenous neural progenitor cells after spinal cord injury in adult rats. Methods: the rat model of acute spinal cord injury was established. Healthy adult rats were divided into normal group, pseudo-operation group and spinal cord injury time group. 5-bromo-2-deoxyuridine (BrdU) tracer, glial fibrillar acidic protein (glial fibrillary acidic protein,GFAP), myelin basic protein (myelin basic protein,MBP) were used to label neural precursor cells, astrocytes and oligodendrocytes, respectively. The number of BrdU labeled positive cells and the number of BrdU/GFAP, BrdU/MBP double labeled positive cells in different groups were observed by immunofluorescence technique, and the proliferation and differentiation of neural progenitor cells after spinal cord injury were investigated. In addition, real-time fluorescence quantitative PCR and Western blotting techniques were used to detect the changing trend of SHH and Gli-1 signal molecules in SHH signaling pathway in rats, and to explore the regulatory effect of SHH signaling pathway after spinal cord injury in rats. Results: a small number of BrdU positive cells and BrdU/GFAP,BrdU/MBP double labeled positive cells were observed in the spinal cord of normal group and false operation group, while the expression of BrdU positive cells and double labeled positive cells increased significantly after spinal cord injury (P 0.05). In addition, the study of SHH signaling pathway components found that the low expression of SHH and Gli-1 signal molecules in the spinal cord of normal rats, the expression of SHH and Gli-1 continued to increase after spinal cord injury, and reached the peak on the 7th day after injury, and remained at a high level on the 21st day after spinal cord injury (P 0.05). Conclusion: the proliferation of neural progenitor cells in spinal cord injury in rats is significantly increased and differentiated into astrocytes and oligodendrocytes. The expression of SHH signal transduction pathway components could be observed in healthy rats. The expression of SHH and Gli-1 protein increased significantly after spinal cord injury, which was basically consistent with the proliferation of neural progenitor cells, suggesting that SHH signaling pathway may be involved in the regulation of nerve cell repair after spinal cord injury.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
本文编号：2504595
[Abstract]:Aim: to observe the proliferation and differentiation of intrinsic neural progenitor cells in spinal cord after acute spinal cord injury, to observe the expression of SHH and Gli-1, which are related to the growth and development of neural progenitor cells, and to explore the molecular mechanism of differentiation regulation of endogenous neural progenitor cells after spinal cord injury in adult rats. Methods: the rat model of acute spinal cord injury was established. Healthy adult rats were divided into normal group, pseudo-operation group and spinal cord injury time group. 5-bromo-2-deoxyuridine (BrdU) tracer, glial fibrillar acidic protein (glial fibrillary acidic protein,GFAP), myelin basic protein (myelin basic protein,MBP) were used to label neural precursor cells, astrocytes and oligodendrocytes, respectively. The number of BrdU labeled positive cells and the number of BrdU/GFAP, BrdU/MBP double labeled positive cells in different groups were observed by immunofluorescence technique, and the proliferation and differentiation of neural progenitor cells after spinal cord injury were investigated. In addition, real-time fluorescence quantitative PCR and Western blotting techniques were used to detect the changing trend of SHH and Gli-1 signal molecules in SHH signaling pathway in rats, and to explore the regulatory effect of SHH signaling pathway after spinal cord injury in rats. Results: a small number of BrdU positive cells and BrdU/GFAP,BrdU/MBP double labeled positive cells were observed in the spinal cord of normal group and false operation group, while the expression of BrdU positive cells and double labeled positive cells increased significantly after spinal cord injury (P 0.05). In addition, the study of SHH signaling pathway components found that the low expression of SHH and Gli-1 signal molecules in the spinal cord of normal rats, the expression of SHH and Gli-1 continued to increase after spinal cord injury, and reached the peak on the 7th day after injury, and remained at a high level on the 21st day after spinal cord injury (P 0.05). Conclusion: the proliferation of neural progenitor cells in spinal cord injury in rats is significantly increased and differentiated into astrocytes and oligodendrocytes. The expression of SHH signal transduction pathway components could be observed in healthy rats. The expression of SHH and Gli-1 protein increased significantly after spinal cord injury, which was basically consistent with the proliferation of neural progenitor cells, suggesting that SHH signaling pathway may be involved in the regulation of nerve cell repair after spinal cord injury.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
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