锌对疼痛后学习记忆障碍小鼠海马神经元P53蛋白表达的影响

doi:10.16098/j.issn.0529-1356.2015.04.004

摘要/Abstract

摘要：

目的探讨锌对第5腰椎（L5）脊神经结扎横切所致神经病理性疼痛（NPP）模型的学习记忆功能障碍及海马组织中P53表达的影响。方法成年小鼠（C57/BL6）48只, 随机分为3组(n=16 )，对照组（con），NPP组及高锌NPP组（Zn-NPP）。对照组仅暴露L5脊神经不结扎，NPP组施行L5脊神经左侧结扎横切（L L5-SNT）手术,Zn-NPP组施行左侧L5-SNT手术后饮用硫酸锌水溶液。3组术后第29 天进行Morris水迷宫实验测量小鼠空间学习记忆能力，水迷宫实验结束后,采用流式细胞术检测各组细胞凋亡率，采用免疫组织化学及免疫印迹法检测各组P53表达。结果 Morris水迷宫实验中,NPP组 (21.03±12.08)m/s，Zn-NPP组(16.97±8.59)m/s及con组(8.74±3.61)m/s，隐藏平台潜伏期逐渐缩短(P< 0.05), 并穿过平台所在象限的时间及路径占总游泳时间和总路径的比例逐渐增加(P< 0.05)。流式细胞术检测结果显示，con组（1.88%），Zn-NPP（16.12%）及NPP组（33.17%）凋亡率逐渐增多(P< 0.05)。免疫组织化学结果显示，con组，Zn-NPP组及NPP组的P53阳性表达吸光度值在CA1区分别为 27.42±7.08、37.52±10.58及53.31±9.26；CA3区分别为29.51±8.31、36.21±7.91及56.71±8.09。免疫印迹结果显示，con组，Zn-NPP组及NPP组的P53表达吸光度值分别为 0.12±1.01、0.37±1.13和0.86±1.22，吸光度值逐渐增多(P< 0.05)。结论锌可改善NPP后小鼠学习记忆能力。

关键词: 锌, 神经病理性疼痛, 学习记忆, P53, 免疫组织化学, 海马, 小鼠

Abstract:

Objective To investigate the effects of the zinc on learning,memory and the expression of P53 in the hippocampus of mice with L5 spinal nerve transection -induced neuropathic pain(NPP). Methods Forty-eight adult male mice were randomly divided into 3 groups (n=16 per group): control group, NPP group and high zinc NPP group (zinc-NPP group). In the control group, the L5 spinal nerve was exposed but without ligation. In the NPP group, the nerve was ligated on the left. The mice of the zinc - NPP group drank a zinc sulfate solution after the left L5 spinal nerve ligation. After 29 days, the spatial learning and memory ability in three groups mice were measured by Morris water maze experiment. After the experiment was finished, cell apoptosis rate was detected by using flow cytometry，P53 expression of each group was tested by immunohistochemical and immune imprinting technology. Results The incubation period of finding the hidden platform was gradually shortened in the NPP group（21.03±12.08）,zinc-NPP group（16.97±8.59）and control group（8.74±3.61) (P<0.05). The proportion of the time and the path to the total time and the total path of finding the Hidden platform gradually increased in NPP group, Zn-NPP, and control group (P<0.05). Flow cytometry detection results shown that the apoptosis rate of Control group（1.88%），Zn-NPP group（16.12%）and NPP group（33.17%）gradually increase(P<0.05).The expression of P53 in the hippocampus CA1 area of the control group，Zn-NPP group and NPP group was 27.42±7.08, 37.52±10.58, and 53.31±9.26, respectively.The expression of P53 in hippocampus CA3 area of the control group，Zn-NPP group and NPP group was 29.51±8.31, 36.21±7.91, and 56.71±8.09, respectively. The expression of P53 in the hippocampus of the control group，Zn-NPP group and NPP group by immune imprinting technology was 29.51±8.31, 36.21±7.91, 56.71±8.09, respectively. All the result showed a gradually increasing trend. Conclusion Zinc can improve the ability of learning and memory in neuropathy pain mice.

Key words: Zinc, Neuropathy pain, Learning and memory, P53, Immunohistochemistry, Hippocampus, Mouse

王月静张海燕廉洁姚宏波孙丽慧* 张萌. 锌对疼痛后学习记忆障碍小鼠海马神经元P53蛋白表达的影响[J]. 解剖学报, 2015, 46(4): 455-461.

WANG Yue-jing ZHANG Hai-yan LIAN Jie YAO Hong-bo SUN Li-hui* ZHANG Meng. Effects of zinc on P53 protein expression in hippocampal neurons of mice with spinal nerve transection -induced neuropathic pain[J]. AAS, 2015, 46(4): 455-461.

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