摘要
目的观察大鼠急性脊髓损伤后不同时间点血管内皮生长因子(VEGF)的表达及局部脊髓组织细胞凋亡的情况,进一步了解脊髓损伤后的病理变化情况。方法雌性SD大鼠,随机分为对照组(A组)、脊髓损伤组(B组),每组各30只。所有大鼠行T12节段椎板切除开窗,暴露脊髓。A组只打开椎板,不打击脊髓。B组按改良Allen法(以10 g×6 cm力撞击脊髓)制作大鼠急性脊髓损伤模型。各组分别于伤后1、3、5、7、14、28 d随机处死5只,损伤部位取材,VEGF、细胞凋亡免疫组化染色及在光镜下观察,计算VEGF阳性细胞数及凋亡细胞数。结果 A组脊髓中央管周围和脊髓软脊膜见少量VEGF表达,其它部位未见表达;B组脊髓损伤后VEGF在脊髓损伤及损伤周边区高表达,5 d达高峰,7 d和14 d表达仍较明显,28 d见少量表达;B组各时间点VEGF表达和A组相比差异均有统计学意义(P<0.05)。A组各时间点凋亡细胞少见,B组则较明显,凋亡细胞在脊髓损伤后逐渐增多,1 d达高峰,3 d和5 d仍较多,7、14、28 d仍有表达;B组各时间点细胞凋亡和A组相比差异均有统计学意义(P<0.05)。结论 VEGF参与急性脊髓损伤后系列病理改变,且持续时间较长。急性脊髓损伤后较长时间内存在着继发性细胞凋亡现象。
Objective To observe the expression level of vascular endothelial growth factor(VEGF) and cells apoptosis at different time points after rats suffering acute spinal cord injury,further in-depth understanding of the pathological changes after spinal cord injury.Methods Female SD rats,which were randomly divided into control group(A group),spinal cord injury group(B group),with 30 animals in each group.The 60 rats were surgically removed the vertebral lamina of T12 segment to expose the spinal cord.And then in A group,the spinal cord was untouched;in B group,the spinal cord was hit by power of 10 g×6 cm according to the improved Allen′s method to establish rat acute spinal cord injury model,then 5 randomly selected rats were sacrificed at 1,3,5,7,14,28 d after injury,materials were obtained from the injury locations following by immunohistochemical staining(IHC) of cell apoptosis and VEGF,observation and calculation of the VEGF positive cells and the positive apoptosis cells under light microscope.Results Except that a few VEGF expressed around central canal and soft meninges of spinal cord,it seldomly expressed in other locations in sham group;In control group,after spinal cord injury VEGF highly expressed in the injured area and its periphery,which reached the peak at 5 d,decreased after 5 d,obviously expressed at 7,14 d and still expressed at 28 d.The expression of VEGF significantly increased at 1,3,5,7,14,28 d compared to control group,and the difference was significant(P<0.05).A little TUNEL labeled positive cells were observed at every time point in sham group,while those cells could be observed obviously after spinal cord injury in the control and intervention groups.Apoptotic cells gradually increased after injury,reached the peak at 1 d,decreased after that.At 5,7 d time points there were still many apoptotic cells and still existed at 14,28 d.The apoptotic cells increased at 1,3,5,7,14,28 d compared to control group,and the difference was significant(P<0.05).Conclusions VEGF plays a part in the pathological change after spinal cord injury,and it persists a long time.Secondary neuron apoptosis exist for a long time after SCI.
出处
《临床骨科杂志》
2013年第1期79-81,共3页
Journal of Clinical Orthopaedics
关键词
脊髓损伤
血管内皮生长因子
细胞凋亡
spinal cord injury
vascular endothelial growth factor
apoptosis
