脊髓横断损伤后再生基因蛋白的表达

目的探讨再生基因蛋白(Reg-2)在脊髓横断损伤后表达的时序变化及其可能的意义。方法SD雌性大鼠45只,实验分为脊髓损伤组(30只)、假手术组(15只)。脊髓损伤组大鼠麻醉后于T9、T10脊髓节段之间用手术刀完全切断脊髓,建立大鼠脊髓横断损伤模型;假手术组大鼠只打开推板,不损伤脊髓,为阴性对照。分别于脊髓损伤后1h、1d、3d、7d1、4d用多聚甲醛灌注后,以T10节段为中心取出长约2cm的脊髓,制作冠状切面及横断切面冷冻切片。分别通过免疫组织化学、免疫荧光和Western blotting法检测Reg-2的表达水平及部位,并与胶质纤维酸性蛋白(GFAP)、少突胶质细胞系转录因子2(Olig2)以及神经肽Y(NPY)、降钙素基因相关肽(CGRP)、神经性生长蛋白-43(GAP-43)等神经细胞特异性标记物做双重标记。结果脊髓损伤组中,表达Reg-2的阳性细胞随着时间的推移,先增加后减少,脊髓损伤后的1周内维持在较高水平,其中第3d后角神经元的Reg-2表达最高,第7d前角神经元Reg-2的表达最高。结论Reg-2可能参与了脊髓损伤后早期的神经再生和重建进程。

THE EXPRESSION OF REGENERATION GENE PROTEIN-2 IN THE SPINAL CORD TRAVERSE INJURY MODEL

Objective To investigate the expression of regeneration gene protein-2(Reg-2) after the transection injury in rat spinal cord. Methods Forty five SD rats were randomly divided into two groups: in the injury group,the SCI was produced by complete transection of the cord at the 9(th) thoracic level,and the sham operation rats were taken as the normal control.The animals were perfused at 1h,1d,3d,7d,14d after the operation,and the spinal cord was taken out at different time points.The expressions of Reg-2 were tested by immunohistochemical analysis and Western blotting,the double staining of Reg-2 and different neural cells special markers(GFAP,Olig2,NPY,CGRP,GAP-43,et al)was done with immunofluorescence method. Results The expression of Reg-2 was found at 1h after the spinal cord injury,reached the peak on day 3 after the injury in the neurons of dorsal horn and day 7 in the neurons of ventral horn.The high expression persisted for 1 week,then decreased gradually.Conclusion Reg-2 may work as an important growth factor and participate in the regeneration and rehabilitation process after the spinal cord injury.