脊髓缺血再灌注损伤后神经元凋亡与热休克蛋白 全部还是部分影响  

Inhibitory effect of heat shock protein on neuronal apoptosis after spinal ischemia-reperfusion injury Thoroughly or partially

作　　者：管华清[1] 杨惠林[1] 徐浩[1] 唐天驷[1] 

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215006

出　　处：《中国临床康复》2005年第25期47-49,i0001,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:诱导抗细胞凋亡的热休克蛋白大量表达,观察其对兔热休克模型脊髓缺血再灌注损伤神经元凋亡的影响。方法:实验于2004-06/09在苏州大学附属第一医院骨科实验室完成。6～8个月新西兰大白兔66只,随机分为3组:①热休克组30只,先制成兔热休克模型,24h后再按脊髓缺血再灌注模型操作。②缺血组30只,单纯制成脊髓缺血再灌注模型。③假手术组6只。除不夹闭腹主动脉外,其余操作同缺血组。④实验过程:于再灌注后8,16,24,48,72h5个时间点热休克组及缺血组各随机抽取6只兔(假手术组6只于8h取出)后肢运动功能进行评分(0分为全瘫,5分为正常)采用Jacob法。后肢运动功能评分后,处死兔并取L2～5脊髓组织观察热休克蛋白70的表达量行免疫组织化学染色法,观察脊髓神经元凋亡情况采用原位缺口末端标记法染色法。计算脊髓前角神经元凋亡率犤凋亡率=阳性神经元数/(阳性神经元数+阴性神经元数)×100%犦。结果:66只白兔均进入结果分析。①兔后肢运动功能Jacob评分:24,48,72h热休克组明显优于缺血组(t=2.825～2.953,P<0.05)。②兔脊髓组织热休克蛋白表达情况:8,16,24,48h热休克组表达量显著高于缺血组(t=8.337～20.470,P<0.01)。③兔脊髓灰质神经元凋亡结果:8,16,24h热休克组明显少于缺血组犤(19.73±3.71)%比(20.87±3.33)%,(27.29±4.20)%比(36.05±5.20)%,(20.64±4.34)%比(35.28±4.27)%,(t=3.884～5.462,P<0.05)犦。结论:热休克蛋白可以明显抑制脊髓缺血再灌注损伤后的神经元凋亡,保护脊髓功能。同时提示热休克蛋白不能全方位地抑制神经元凋亡,部分神经元的凋亡对兔后肢运动不产生显著影响。AIM： To induce the expression of heat shock protein （HSP） with the function of anti-apoptosis, and observe its inhibitory effect on the apoptosis of spinal neuron after ischemia-reperfusion （IR） injury in rabbit models of heat shock. METHODS： The experiment was conducted in the laboratory of Department of Orthopaedics in the First Affiliated Hospital of Soochow University from June to September 2004. Sixty-six New Zealand white big rabbits at the age of 6 to 8 months were randomly divided into 3 groups： ① heat shock group （n=30）, in which models of rabbit heat shock were established firstly, and 24 hours later they were operated into the spinal IR models. ② ischemia group （n=30）, in which rabbits were established into spinal IR models only, ③ sham operation group（n=6）, in which the rabbits were operated as the IR group were, except for closure by clip of abdominal aorta. At the time of 8, 16, 24, 48 and 72 hours after reperfusion, 6 rabbits of both ischemia and heat shock group were picked out randomly （all the rabbits of sham operation group were picked out at 8 hours）,and then their motor function of posterior limbs was assessed according to Jacob＇s scoring （0 point as paraplegia and 5 points as health）. After they were sacrificed, the spinal cords（L2-L5） were removed. The expression of HSP 70 was examined by immunohistochemical staining, and the apoptosis of nerve cells by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL） staining method. The apoptosis rate of nerve cells in anterior spinal corn was calculated based on the equation： apoptosis rate=number of positive nerve eells/（number of positive nerve cells＋number of negative nerve cells）×100%. RESULTS： All the 66 white rabbits were analyzed in the result. ① The Jacob＇s score of motor function of rabbit posterior limbs in heat shock group was significantly higher than that in the isehemia group at 24, 48 and 72 hours（t=2.825 to 2.953, P 〈 0.05）. ② The expressi

关 键 词：脊髓损伤 缺血 再灌注 热休克蛋白质70 

分 类 号：R744[医药卫生—神经病学与精神病学]