摘要
目的观察大鼠脊髓损伤后低氧诱导因子-1α(HIF-1α)的表达规律,探讨其在脊髓损伤发生发展过程中的作用及法医学应用。方法建立大鼠静压脊髓损伤模型,采用RT-PCR和免疫组化SP法对伤后不同时间(0、3、6、12h和1、3、5、7、11、14d)HIF-1α和HIF-1αmRNA的表达进行检测,BI2000图像分析系统分析结果。结果正常及假手术对照组大鼠脊髓组织内有低水平的HIF-1αmRNA表达,但几乎检测不到HIF-1α阳性细胞;脊髓损伤后,HIF-1α及HIF-1αmRNA表达开始升高,其中HIF-1αmRNA表达在伤后3h开始增加,3d达高峰,14d时恢复正常;HIF-1α表达在伤后3h开始增加,1d达高峰,较HIF-1αmRNA的达峰时间(伤后3d)提前。HIF-1α免疫阳性产物可见于脊髓神经元、胶质细胞、室管膜细胞及间质内的血管内皮细胞。结论脊髓损伤后HIF-1α基因表达开始升高,对组织和细胞起低氧保护作用,其表达的时序性规律可望用于法医学损伤时间推断。
Objective To investigate the expression changes of hypoxia inducible factor-1α （ HIF-1α） in rats after experiment spinal cord injury（SCI）. Methods 66 SD rats were divided into a normal control, a sham operation and 9 different injury groups at 3,6,12h and 1,3,5,7,11,14d after SCI. The static compression model of SCI was employed in the injury groups, and the expression of HIF-1α at levels of protein and mRNA in different wound age were detected by immunohistochemical and RT-PCR techniques. Results In spinal cord of normal control and sham operation groups, the level of HIF-1α mRNA was low and nearly no HIF-1α positive cell could be detected, and no difference was found between these two groups（P 〉0.05 ）. In spinal cord tissues of injury groups, the level of HIF-1α mRNA began to elevate at 3h after injury and reached the maximize on 3d, then it began to decline and returned to the basal level on 14d; the HIF-1α expression showed a time-dependent difference as followed： It began to increase at 3h after injury, peaked on 1d, then gradually decreased, and recovered to the normal level on 14d
afterinjury. Conclusion The result suggested that hypoxia circumstance can enhance the stability of HIF-1α and the transcription of HIF-1α mRNA, and the time-dependent expression of HIF-1α and HIF-1α mRNA can be used to help the diagnosis of SCI and estimation of injury time.
出处
《中国法医学杂志》
CSCD
2008年第1期4-7,共4页
Chinese Journal of Forensic Medicine
