CAJ | 학술논문

目的探讨腺苷处理后犬脊髓缺血再灌注损伤实验中脊髓神经元凋亡及其相关蛋白Bax、Bcl-2的表达变化,证实腺苷对犬脊髓缺血再灌注的保护作用.方法 16只健康成年杂种犬,雌雄不拘,体质量10 ～20 kg.随机均分为两组,采用胸主动脉、锁骨下动脉阻断法造成脊髓缺血模型(75 min).单纯缺血再灌注组,阻断前30 min颈静脉泵入50 ml生理盐水,300 ml/h,10 min泵完;阻断结束时再以相同的流速和时间泵完(对照组).阻断前30 min颈静脉泵入50 ml腺苷溶液(0.4 mg/ml,50 ml),阻断结束时再以相同的流速相同浓度和时间泵完(实验组).术后继续笼内饲养,采用Tarlov下肢神经功能评分系统评估6、24、48 h神经运动功能评分.再灌注48 h各组分别处死实验动物,迅速取出L2 ～L5及骶段脊髓.采用Tunel法进行细胞凋亡检测,免疫组化法检测Bax、Bcl-2的表达.结果实验组犬运动功能评分24、48 h高于对照组,差异有统计学意义(P＜0.05).实验组脊髓神经元凋亡量明显低于对照组(P＜0.05),较对照组Bax表达减少和Bcl-2表达增多.结论腺苷处理后可减少犬脊髓缺血再灌注后神经元凋亡,对脊髓缺血再灌注损伤具有明显的保护作用.其保护作用可能与凋亡蛋白Bax表达减少和凋亡抑制蛋白Bcl-2表达增多有关.
목적 탐토선감처리후견척수결혈재관주손상실험중척수신경원조망급기상관단백Bax、Bcl-2적표체변화,증실선감대견척수결혈재관주적보호작용.방법 16지건강성년잡충견,자웅불구,체질량10 ～20 kg.수궤균분위량조,채용흉주동맥、쇄골하동맥조단법조성척수결혈모형(75 min).단순결혈재관주조,조단전30 min경정맥빙입50 ml생리염수,300 ml/h,10 min빙완;조단결속시재이상동적류속화시간빙완(대조조).조단전30 min경정맥빙입50 ml선감용액(0.4 mg/ml,50 ml),조단결속시재이상동적류속상동농도화시간빙완(실험조).술후계속롱내사양,채용Tarlov하지신경공능평분계통평고6、24、48 h신경운동공능평분.재관주48 h각조분별처사실험동물,신속취출L2 ～L5급저단척수.채용Tunel법진행세포조망검측,면역조화법검측Bax、Bcl-2적표체.결과 실험조견운동공능평분24、48 h고우대조조,차이유통계학의의(P＜0.05).실험조척수신경원조망량명현저우대조조(P＜0.05),교대조조Bax표체감소화Bcl-2표체증다.결론 선감처리후가감소견척수결혈재관주후신경원조망,대척수결혈재관주손상구유명현적보호작용.기보호작용가능여조망단백Bax표체감소화조망억제단백Bcl-2표체증다유관.
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.