活血通督汤对脊髓缺血再灌注损伤神经细胞凋亡的影响

目的:通过观察活血通督汤对脊髓缺血再灌注损伤脊髓神经细胞凋亡的影响,探讨活血通督汤改善脊髓缺血再灌注损伤的机制。方法:66只新西兰家兔随机分为假手术组、模型组、活血通督汤组。模型组、活血通督汤组采用夹闭腰动脉法制作脊髓缺血再灌注损伤模型。假手术组不夹闭腰动脉,其它步骤同模型组。采用TUNEL法检测神经细胞凋亡水平。结果:(1)苏木精-伊红染色显示:假手术组脊髓神经细胞形态基本正常;模型组和活血通督汤组各时间点可见脊髓神经细胞核固缩、深染、结构模糊,间质可见出血点;活血通督汤组各时间点脊髓神经细胞形态优于模型组。(2)TUNEL染色显示:假手术组神经细胞凋亡较少;模型组和活血通督汤组各时间点神经细胞凋亡较假手术组明显增高,再灌注24h神经细胞凋亡最多;活血通督汤组各时间点脊髓神经细胞凋亡少于模型组。结论:活血通督汤可通过减少脊髓缺血再灌注损伤中脊髓神经细胞的凋亡,从而减轻脊髓缺血再灌注损伤。

Study on the Apoptosis of Neurons during Spinal Cord Ischemic-reperfusion Injury Treated by Huoxue Tongdu Decoction

Objective：To investigate the effect of Huoxue Tongdu Decoction on apoptosis of neurons during spinal cord is- chemie-reperfusion injury, and to explore the underlying mechanism. Methods： New Zealand rabbits were randomly divided into three groups： sham-operated group （n=6）, model group （n： 30）, and Huoxue Tongdu Decoction group （n= 30）. Lumbar artery occlusion from the third to the sixth vertebrae was performed to induce spinal cord ischemia in the model and Huoxue Tongdu Decoction groups, while lumbar artery occlusion was not performed in the sham operation group. Blood flow was recovered 30 minutes following occlusion to establish models of spinal cord ischemia-reperfusion injury. The apop- tosis of spinal cord neurons were observed by HE-staining and TUNEL labeling at the time points of 8h, 12h, 24h, 3d, and lw following perfusion. Results： （1） HE：-staining showed that he shape and structure of the neurons was normal in the sham-operation group. Compared with sham operation group, neurons showed obvious degeneration at different time points in the model and Huoxue Tongdu Decoction groups. The shape and structure of the neurons in Huoxue Tongdu Decoction group were better than the model groups. （2） TUNEL labeling showed that the apoptotic neurons were lowest in the sham- operation group. Compared with the sham operation group, apoptotic neurons obviously increased at different time points in the model and Huoxue Tongdu Decoction groups （P〈0.01）. The apoptotic neurons in Huoxue Tongdu Decoction group were significantly decreased compared with the model group （P〈0.01）. Conclusion.. Huoxue Tongdu Decoction can im- prove spinal cord ischemic-reperfusion injury by reducing the apoptosis of spinal cord neurons.