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

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∗
苏建华;陈玉芳;唐金荣;丁新生;肖杭
【期刊名称】《医药导报》
【年(卷),期】2016(35)6
【摘要】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 and 100 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,
pared 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 24th hours&nbsp;(P<0.01).Compared with low-dose methycobal group, there were significant differences in the high-dose methycobal group at the end of 6th, 12th and 24th hours(P<0.01 or P<0.05).The results of RT-PCR were as follows: there was expression of caspase-3 mRNA in the perifocal cortex of all groups, with weak expression in the sham-operation pared with the sham-operation group, the expression of caspase-3 mRNA was increased significantly in the model control group(P<0.01).The expression of caspase-3 mRNA was reduced significantly in the nimodipine group, the low-dose methycobal group and high-dose methycobal group as compared with model control group at each time point( P<0.05 or P<0.01) , but it was not significantly different in the low-dose methycobal group and high-dose methycobal group as compared with that of the nimodipine group(P>0.05).There were
significant differences between the high-dose methycobal group and low-dose methycobal group at the end of 24 h(P<0.05). Conclusion Methycobal can protect the brain cells from injury after cerebral ischemia reperfusion by adjusting the expression of Caspase-3m RNA, and the high-dose methycobal is more effective.%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 mL.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。

【总页数】5页(P574-578)
【作者】苏建华;陈玉芳;唐金荣;丁新生;肖杭
【作者单位】江苏大学附属金坛医院神经内科，金坛 213200;江苏省常州市第四
人民医院病理科，常州 213000;南京医科大学第一附属医院神经内科，南京210029;南京医科大学第一附属医院神经内科，南京 210029;南京医科大学应用神经毒理研究所，南京 210029
【正文语种】中文
【中图分类】R972;R961
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