摘要
目的探究大鼠脊髓损伤后磷脂酰乙醇胺结合蛋白1(PEBP1)的表达和意义。方法本研究采用105只Sprague-Dawley雌性大鼠,随机分为假手术组和实验组。采用Allen’s法制备大鼠脊髓损伤模型,应用免疫组织化学方法检测PEBP1在脊髓中的定位变达,采用实时定量PCR和蛋白免疫印迹检测PEBP1的mRNA和蛋白在脊髓中的表达变化。结果免疫组织化学结果显示PEBP1在正常脊髓后角的神经胶质细胞的胞浆和胞核中表达,在脊髓损伤后与sham组相比PEBP1免疫阳性细胞数明显减少。此外,实时定量PCR显示PEBP1的mRNA在大鼠脊髓损伤后12h,1d,3d,7d,14d和28d的表达量明显降低,与sham组相比差异具有统计学意义(P<0.01);免疫印迹实验显示,与sham组相比,PEBP1蛋白在大鼠脊髓损伤后的7d,14d和28d表达量降低,差异具有统计学意义(P<0.05)。结论脊髓损伤后PEBP1的mRNA和蛋白表达水平均降低,在脊髓后角的表达明显减少。提示脊髓损伤后PEBP1可能参与调控神经胶质细胞的存活和凋亡,进一步影响脊髓损伤后的感觉功能。
Objective This study aimed to investigate the expression and significance of phosphatidylethanolaminebinding protein 1（PEBP1）following acute spinal cord injury in rats.Methods One hundred and five adult female Sprague-Dawley rats were used and randomly divided into two groups which contain sham group and spinal cord injury group.Rat model of contusion spinal cord injury was established with modified Allen＇s method.Immunohistochemistry staining was used to determined the distribution of PEBP1 in spinal cord.Furthermore,real-time quantitative polymerase chain reaction（qRT-PCR）and western blot were used to detected the level of PEBP1 mRNA and protein in spinal cord.Results Immunohistochemistry staining showed PEBP1 was found in the posterior horn of spinal cord,locating in the cytoplasm and nucleus of glia,and the number of positive cells decreased significantly after spinal cord injury when compared to sham group.In addition,qRT-PCR showed that the level of PEBP1 mRNA decreased significantly at 12 h,1d,3d,7d,14 dand 28dafter spinal cord injury（P〈0.01）.The data of western blot showed that the level of PEBP1 protein reduced significantly at 7d,14 dand 28d（P〈0.05）.Conclusion This study found that the level of PEBP1 mRNA and protein reduced in rats with spinal cord injury,and the expression of PEBP1 in posterior horn was weaken after spinal cord injury.Results indicated that PEBP1 might be associated with the glia survival and apoptosis,which can effect of the sensory function after spinal cord injury.
出处
《四川解剖学杂志》
2015年第4期1-6,共6页
Sichuan Journal of Anatomy
基金
国家级大学生创新实验训练计划(201313705005)
大学生创新实验项目(CXXS201306)
