The neuronal apoptosis and related gene expression changes after adenosine treatment in canine spinal cord ischemiareperfusion injury experiments

Abstract

Objective To investigate the protective effect of adenosine on spinal cord Ischemia/Reperfusion through the observation of the neurons'apoptosis and the change of Bax and Bcl-2.Methods 16 healthy adult mongrel canines of either sex,weighted 10-20kg,were randomly divided into two groups (n =8).Creating the models of spinal cord Ischemia/Reperfusion by thoracic aorta and subclavian artery occlusion.Control group:i.e the simple Ischemia/Reperfusion group.Pumping 50 ml saline through the jugular vein 30 minutes before blocking the artery,and the rate is 300ml / h,pumping time 10 minutes.Then pump another 50ml saline right after blocking at the same rate.Experimental group:20mg adenosine were dissolved into 50ml saline and was pumped 30min before blocking the artery in the same way.Still pump another 50ml adenosine solution of the same concentration right after blocking.After the operation,canines were continued to feed with free access to food and water.Recording the motor function score at 6 h,24 h,48 h after reperfusion (see Taylor motor function scoring system).All 8 canines were killed after 48 hours of reperfusion.L2-L5 and the sacral spinal cord were taken out as quickly as possible.The Tunel method was adopted to detect neurons'apoptosis,and the immunohistochemical method to detect the expression changes of Bax and Bcl-2.Results In the experimental group,the 24 h,48 h motor function scores were significantly higher than that in the control group (P ＜ 0.05).The neurons'apoptosis was significantly lower than that of the control group (P ＜ 0.05).Compared with the control group,the expression of the apoptotic protein Bax reduced while the apoptosis-inhibited protein Bcl-2 increased.Conclusion The neurons'apoptosis can be reduced significantly through the adenosine treatment and adenosine has a apparently protective effect on canines'spinal cord Ischemia/Reperfusion injury.The effect might be related to Bax and Bcl-2. Key words: Adenosine; Neurons' apoptosis; Apoptosis-related protein ; Spinal cord Ischemia/Reperfusion injury; Protection