甲钴胺对大鼠缺血-再灌注损伤神经细胞凋亡及Caspase-3 mRNA表达的影响  

Effect of Methycobal on Neuron Apoptosis of Caspase-3 mRNA Expression in Rat Brain Tissue After Cerebral Ischemia Reperfusion

作　　者：苏建华[1] 陈玉芳[2] 唐金荣[3] 丁新生[3] 肖杭[4] 

机构地区：[1]江苏大学附属金坛医院神经内科,金坛213200 [2]江苏省常州市第四人民医院病理科,常州213000 [3]南京医科大学第一附属医院神经内科,南京210029 [4]南京医科大学应用神经毒理研究所,南京210029

出　　处：《医药导报》2016年第6期574-578,共5页Herald of Medicine

基　　金：江苏省"333高层次人才培养工程"科研项目资助(BRA2012050);江苏大学医学临床科技发展基金资助项目(JLY2010053);常州市科技局资助项目(WZ201043)

摘　　要：目的探讨甲钴胺对缺血-再灌注大鼠脑组织半胱氨酸天冬氨酸酶3(Caspase-3)mRNA表达的影响。方法将大鼠按随机原则分为假手术组、模型对照组、尼莫地平组、甲钴胺小剂量组、甲钴胺大剂量组,每组30只。假手术组与模型对照组分别灌胃给予0.9%氯化钠溶液20 m L·kg^(-1)·d^(-1);尼莫地平组灌胃尼莫地平1 mg·kg^(-1)·d^(-1);甲钴胺小、大剂量组分别灌胃甲钴胺50,100μg·kg^(-1)·d^(-1),给药1周。采用线栓法栓塞大脑中动脉3 h制作大鼠脑缺血-再灌注模型;观察再灌注24 h后神经功能缺损评分,于6,12,24 h用TUNEL法检测顶叶皮质细胞凋亡、反转录聚合酶链反应(RT-PCR)检测Caspase-3 mRNA的表达。结果模型对照组神经功能缺损评分为(2.70±0.52)分,尼莫地平组、甲钴胺小剂量组、甲钴胺大剂量组分别为(1.30±0.51),(2.20±0.75),(1.30±0.81)分,与模型对照组比较,均差异有统计学意义(P<0.05或P<0.01);与尼莫地平组比较,甲钴胺大剂量组差异无统计学意义(P>0.05),与甲钴胺小剂量组比较,甲钴胺大剂量组差异有统计学意义(P<0.05)。尼莫地平组、甲钴胺小、大剂量组各时间点神经细胞凋亡较模型对照组明显减少;甲钴胺大剂量组24 h时较尼莫地平组差异有统计学意义(P<0.01);6~24 h较甲钴胺小剂量组差异有统计学意义(P<0.01或P<0.05)。假手术组Caspase-3 mRNA少量表达,与假手术组比较,模型对照组6~24 h Caspase-3 mRNA表达差异有统计学意义(P<0.01);与模型对照组比较,尼莫地平组、甲钴胺大、小剂量组6~24 h Caspase-3 mRNA表达差异有统计学意义(P<0.05或P<0.01);与尼莫地平组比较,甲钴胺大、小剂量组各时间点Caspase-3 mRNA差异无统计学意义(P>0.05),与甲钴胺小剂量组比较,甲钴胺大剂量组于24 h差异有统计学意义(P<0.05)。结论甲钴胺对脑缺血-再灌注损伤保护机制与抑制Caspase-3 mRNA表达有关,且以大剂量效果较好。Objective To investigate the effects of methycobal on the expression of Caspase-3 in brain tissue after cerebral ischemia reperfusion in rats. Methods Rats were randomly divided into sham-operation group,model control group,nimodipine group and low-dose methycobal group,high-dose methycobal group（ n = 30 in each group）.Rats in the sham-operation group and model control group were administered intragastrically with 0.9% sodium chloride solution,rats in the nimodipine group were treated with 1 mg · kg-（-1）·d-（-1）of nimodipine,rats in the low- and high-dose of methycobal groups were given 50 and100 μg·kg-（-1）·d-（-1）of methycobal,respectively. The rat model of cerebral ischemia reperfusion was established by middle cerebral artery occlusion with suture method for 3 h.Neurological deficit scores were evaluated 24 h after reperfusion.The apoptosis of perifocal cortex cells was detected by TUNEL method and the expression of Caspase-3 was analyzed by RT-PCR 6,12 and 24 h after reperfusion. Results Neurological deficit scores in model control group,nimodipine group,low-dose methycobal group and high-dose methycobal group were 2.70 ± 0. 52,1. 30 ± 0. 51,2. 20 ± 0. 75 and 1. 30 ± 0. 81,respectively. Compared with model control group,neurological deficit scores were significantly different in the nimodipine group,low-dose methycobal group and high-dose methycobal group（ P〈0.05 or P〈0.01）. There were no significant differences between the high-dose methycobal group and nimodipine group（ P〈0. 05）. There was a significant difference between the high-dose methycobal group and low-dose methycobal group（ P〈0. 05）. The results of apoptosis by TUNEL were as follows： compared with model control group,the apoptosis decreased obvsiouly in the nimodipine group,low-dose methycobal group,and high-dose methycobal group at each time point.There was significant difference between the high-dose methycobal group and nimodipine group at the end of the 24thhours（ P〈0.01）.Compared with low-

关 键 词：甲钴胺 脑缺血-再灌注 细胞凋亡 半胱氨酸天冬氨酸酶3 

分 类 号：R972[医药卫生—药品]