Noggin蛋白对小鼠脑缺血再灌注损伤后学习和记忆能力与齿状回结构的影响

doi:10.6040/j.issn.1671-7554.0.2019.1273

摘要/Abstract

摘要： 目的探讨侧脑室注射Noggin蛋白对小鼠脑缺血再灌注损伤(CIRI)后学习、记忆能力及齿状回(DG)结构的影响,以期为临床缺血性脑血管疾病的预防与治疗提供新思路。方法 240只小鼠随机分为:假手术组(n=80)、缺血再灌注损伤组(I/R组,n=80)、缺血再灌注损伤+Noggin治疗组(Noggin组,n=80),每组再分为1、3、7、14 d共 4个亚组;取材前1天采用Y型迷宫检测小鼠学习记忆能力(n=5);然后比色法检测缺血侧脑组织超氧化物歧化酶(SOD)与丙二醛(MDA)含量(n=5);2,3,5-氯化三苯基四氮唑(TTC)染色检测脑梗死面积(n=5); 尼氏染色与免疫荧光检测DG神经元与胶质原纤维酸性蛋白(GFAP)阳性细胞变化(n=5);Western blotting法检测骨形态发生蛋白4(BMP4)蛋白表达(n=5)。结果随着损伤时间的增加,I/R组、Noggin组与假手术组相比,小鼠神经功能评分升高(F组别=21.19, P<0.001; F时间=25.13, P<0.001),学习、记忆能力均下降［(F组别=216.10, P<0.001; F时间=260.10, P<0.001)、(F组别=114.40, P<0.001; F时间=184.60, P<0.001)］,脑梗死面积增加(F组别=2374, P<0.001; F时间=3 292, P<0.001),SOD活性降低(F组别=1 426, P<0.001; F时间=1 723, P<0.001),MDA含量升高(F组别=2.22, P<0.001; F时间=6.33, P<0.001),神经元数量减少(F组别=148.90, P<0.001; F时间=485.50, P<0.001),GFAP阳性细胞和BMP4蛋白表达量增加［(F组别=40.18, P<0.001; F时间=141.90, P<0.001)、(F组别=426.70, P<0.001; F时间=1 329, P<0.001)］。在各个相同时间点,与I/R组相比,Noggin各组小鼠神经功能评分降低(P<0.001),学习、记忆能力提高(P<0.001),脑梗死面积减小(P<0.001),SOD活性升高及MDA含量降低(P<0.001),神经元数量增加及染色变浅(P<0.001),GFAP阳性细胞和BMP4 蛋白表达量降低(P<0.001)。结论 Noggin蛋白对小鼠脑缺血再灌注损伤后学习、记忆能力提高与DG组织损伤修复有改善作用,其作用机制可能与阻断BMP4蛋白表达和抑制胶质细胞的反应性增生有关。

关键词: Noggin蛋白, 脑缺血再灌注损伤, 齿状回, 骨形态发生蛋白4, 胶质原纤维酸性蛋白, 小鼠

Abstract: Objective To investigate the effect of injection of Noggin protein into the lateral ventricle on the learning and memory abilities and the structure of dentate gyrus after cerebral ischemia reperfusion injury(CIRI)in mice, in order to provide new ideas for the prevention and treatment of clinical ischemic cerebrovascular diseases. Methods A total of 240 health mice were randomly divided into the sham group(n=80), the ischemia-reperfusion group(I/R group, n=80), and the Noggin treatment group(Noggin group, n=80). Each group was subdivided into the 1days, 3 days, 7 days and 14 days subgroups. The learning and memory abilities were detected by Y-maze at 1 day before execution. Infarct size was detected by triphenyltetrazolium chloride staining(n=5). The colorimetric method was used to detect the superoxide dismutase(SOD)and malondialdehyde(MDA)activity after the behavior capability determined(n=5). Infarct size was detected by 2,3,5-triphenyl tetrazolium chloride staining(n=5). Glial fibrillary acidic protein(GFAP)immunopositive cells in the DG were observed under a light microscopy and quantitative analysis were performed by cell morphometric technique(n=5). The BMP4 protein expression was detected by Western blotting(n=5). Results Compared with the sham group, the neurological function score of mice in the I/R group and Noggin group increased(F_group=21.19, P<0.001; F_time=25.13, P<0.001), learning and memory abilities declined(F_group=216.10, P<0.001; F_time=260.10, P<0.001)(F_group=114.40, P<0.001; F_time=184.60, P<0.001), and area of cerebral infarction increased(F_group=2 374, P<0.001; F_time=3 292, P<0.001), SOD activity reduced(F_group=1 426, P<0.001; F_time=1 723, P<0.001)and MDA content increased(F_group=2.22, P<0.001; F_time=6.33, P<0.001), number of neurons reduced(F_group=148.90, P<0.001; F_time=485.50, P<0.001), and expression of GFAP positive cells and BMP4 protein increased(F_group=40.18, P<0.001; F_time=141.90, P<0.001)(F_group=426.70, P<0.001; F_time=1 329, P<0.001)with the increase of injury time. At each same time point, compared with the I/R group, the Noggin groups neurological function score decreased(P<0.001), learning and memory abilities increased(P<0.001), and cerebral infarction area decreased(P<0.001), SOD activity increased and MDA content decreased(P<0.001), number of neurons increased and staining was lightened (P<0.001), expression of GFAP positive cells and BMP4 protein decreased(P<0.001). Conclusion Noggin can efficiently improve the learning and memory abilities and decrease the pathological lesion of DG after cerebral ischemia reperfusion injury in mice, and its mechanism may closely related to BMP4 protein expression and glial cells activation.

Key words: Noggin protein, Cerebral ischemia reperfusion injury, Dentate gyrus, Bone morphogenetic protein 4, Glial fibrillary acidic protein, Mouse

中图分类号:

R743.31

龙婷婷,谢明,周璐,朱俊德. Noggin蛋白对小鼠脑缺血再灌注损伤后学习和记忆能力与齿状回结构的影响[J]. 山东大学学报 (医学版), 2020, 58(7): 15-23.

LONG Tingting, XIE Ming, ZHOU Lu, ZHU Junde. Effect of Noggin protein on learning and memory abilities and the dentate gyrus structure after cerebral ischemia reperfusion injury in mice[J]. Journal of Shandong University (Health Sciences), 2020, 58(7): 15-23.

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