您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报 (医学版) ›› 2020, Vol. 1 ›› Issue (7): 15-23.doi: 10.6040/j.issn.1671-7554.0.2019.1273

• • 上一篇    下一篇

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

龙婷婷,谢明,周璐,朱俊德   

  1. 贵州医科大学基础医学院人体解剖学教研室, 贵州 贵安 550025
  • 出版日期:2020-07-20 发布日期:2020-07-10
  • 通讯作者: 朱俊德. E-mail:jdzhu73@126.com
  • 基金资助:
    国家自然科学基金(81660243);贵州省科技厅社会发展科技计划(黔科合SY字［2015］3041号)

Effect of Noggin protein on learning and memory abilities and the dentate gyrus structure after cerebral ischemia reperfusion injury in mice

LONG Tingting, XIE Ming, ZHOU Lu, ZHU Junde   

  1. Department of Human Anatomy, School of Basic Medical Science, Guizhou Medical University, Guian 550025, Guizhou, China
  • Online:2020-07-20 Published:2020-07-10

PDF (PC)

286

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, zw

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(Fgroup=21.19, P<0.001; Ftime=25.13, P<0.001), learning and memory abilities declined(Fgroup=216.10, P<0.001; Ftime=260.10, P<0.001)(Fgroup=114.40, P<0.001; Ftime=184.60, P<0.001), and area of cerebral infarction increased(Fgroup=2 374, P<0.001; Ftime=3 292, P<0.001), SOD activity reduced(Fgroup=1 426, P<0.001; Ftime=1 723, P<0.001)and MDA content increased(Fgroup=2.22, P<0.001; Ftime=6.33, P<0.001), number of neurons reduced(Fgroup=148.90, P<0.001; Ftime=485.50, P<0.001), and expression of GFAP positive cells and BMP4 protein increased(Fgroup=40.18, P<0.001; Ftime=141.90, P<0.001)(Fgroup=426.70, P<0.001; Ftime=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, zw

中图分类号: 

  • R743.31
[1] Lu H, Liu X, Zhang N, et al. Neuroprotective effects of brain-derived neurotrophic factor and noggin-modified bone mesenchymal stem cells in focal cerebral ischemia in rats[J]. J Stroke Cerebrovasc Dis, 2016, 25(2): 410-418.
[2] Xu H, Liu W, Liu T, et al. Synergistic neuroprotective effects of Danshensu and hydroxysafflor yellow A on cerebral ischemia-reperfusion injury in rats[J]. Oncotarget, 2017, 8(70): 115434-115443.
[3] Wang C, Jiang J, Zhang X, et al. Inhibiting HMGB1 reduces cerebral ischemia reperfusion injury in diabetic mice[J]. Inflammation, 2016, 39(6): 1862-1870.
[4] Young JJ, Kjolby RAS, Wu G, et al. Noggin is required for first pharyngeal arch differentiation in the frog Xenopus tropicalis[J]. Dev Biol, 2017, 426(2): 245-254.
[5] Smith WC, Harland RM. Expression cloning of noggin, a new dorsalizing factor localized to the Spemann organizer in Xenopus embryos[J]. Cell, 1992, 70(5): 829-840.
[6] 李伟. 阿尔茨海默病的影响因素和生物标志物研究[D]. 上海:上海交通大学,2017.
[7] Díaz-Moreno M, Armenteros T, Gradari S, et al. Noggin rescues age-related stem cell loss in the brain of senescent mice with neurodegenerative pathology[J]. Proc Natl Acad Sci U S A, 2018, 115(45): 11625-11630.
[8] Zimmerman LB, De Jesús-Escobar JM, Harland RM, et al. The Spemann organizer signal noggin binds and inactivates bone morphogenetic protein 4[J]. Cell, 1996, 86(4): 599-606.
[9] 王俊婕,张先虎,朱俊德,等. DAPT对脑缺血再灌注损伤小鼠学习记忆的影响以及Notch信号通路的调节作用[J].解剖学报,2018,49(2):151-157. WANG Jun Jie, ZHANG Xian Hu, ZHU Jun De, et al. Effects of DAPT on learning and memory abilities and the regulation of Notch signaling pathway in cerebral ischemia-reperfusion injury mice[J]. Acta Anatomica Sinica, 2018, 49(2): 151-157.
[10] Mikawa S, Sato K. Noggin expression in the adult rat brain[J]. Neuroscience, 2011, 184: 38-53. doi: 10.1016/j.neuroscience.2011.03.036.
[11] Zhu JD, Wang JJ, Ge G, et al. Effects of noggin-transfected neural stem cells on neural functional recovery and underlying mechanism in rats with cerebral ischemia reperfusion injury[J]. J Stroke Cerebrovasc Dis, 2017, 26(7): 1547-1559.
[12] Shin JA, Kim YA, Kim HW, et al. Iron released from reactive microglia by noggin improves myelin repair in the ischemic brain[J]. Neuropharmacology, 2018, 133: 202-215. doi: 10.1016/j.neuropharm.2018.01.038.
[13] Wang JJ, Zhu JD, Zhang XH, et al. Neuroprotective effect of Notch pathway inhibitor DAPT against focal cerebral ischemia/reperfusion 3 hours before model establishment[J]. Neural Regen Res, 2019, 14(3): 452-461.
[14] Li S, Bian L, Fu X, et al. Gastrodin pretreatment alleviates rat brain injury caused by cerebral ischemic-reperfusion[J]. Brain Res, 2019, 1712: 207-216. doi: 10.1016/j.brainres.2019.02.006.
[15] Su Q, Zhang H, Dang S, et al. Hippocampal protein kinase C gamma signaling mediates the impairment of spatial learning and memory in prenatally stressed offspring rats[J]. Neuroscience, 2019, 414: 186-199. doi: 10.1016/j.neuroscience.2019.06.030.
[16] Bak J, Pyeon HI, Seok JI, et al. Effect of rotation preference on spontaneous alternation behavior on Y maze and introduction of a new analytical method, entropy of spontaneous alternation[J]. Behav Brain Res, 2017, 320: 219-224. doi: 10.1016/j.bbr.2016.12.011.
[17] He F, Dai R, Zhou X, et al. Protective effect of 4-Methoxy benzyl alcohol on the neurovascular unit after cerebral ischemia reperfusion injury[J]. Biomed Pharmacother, 2019, 118: 109260. doi: 10.1016/j.biopha.2019.109260.
[18] Zhang B, Zhang HX, Shi ST, et al. Interleukin-11 treatment protected against cerebral ischemia/reperfusion injury[J]. Biomed Pharmacother, 2019, 115: 108816. doi: 10.1016/j.biopha.2019.108816.
[19] Meyers EA, Gobeske KT, Bond AM, et al. Increased bone morphogenetic protein signaling contributes to age-related declines in neurogenesis and cognition[J]. Neurobiol Aging, 2016, 38: 164-175. doi: 10.1016/j.neurobiolaging.2015.10.035.
[20] 朱俊德,余资江,康朝胜,等. noggin蛋白对D-半乳糖致衰老小鼠学习记忆及海马齿状回神经发生的影响[J].山东大学学报(医学版),2009,30(8):72-74. ZHU Jun De, YU Zi Jiang, KANG Chao Sheng, et al. Effect of intracerebroventricular injection of noggin on learning and memory and neurogenesis in hippocampal dentate gyrus of D-galactose induced aging mice[J]. Journal of Shandong University(Health Sciences), 2009, 30(8): 72-74.
[21] 吴嶛,孙颖,孙波,等. 辛醇对大鼠脑缺血再灌注后星形胶质细胞和胶质原纤维酸性蛋白的影响[J].中国医药,2019,14(5):706-709. WU Liao, SUN Ying, SUN Bo, et al. Effect of octanol on astrocytes and glial fibrillary acidic protein in rats after cerebral ischemia-reperfusion[J]. China Medicine, 2019, 14(5): 706-709.
[22] 欧阳波,陈勇军,唐小容,等. 小RAN干扰Trx1促进大鼠缺血再灌注损伤的星形胶质细胞凋亡、抑制其增殖及作用机制[J].中国老年学杂志,2019,39(9):192-195.
[23] Watanabe T, Nagai A, Sheikh AM, et al. A human neural stem cell line provides neuroprotection and improves neurological performance by early intervention of neuroinflammatory system[J]. Brain Res, 2016, 1631: 194-203. doi: 10.1016/j.brainres.2015.11.031.
[24] Hamada-Kanazawa M, Ogawa D, Takano M, et al. Sox6 suppression induces RA-dependent apoptosis mediated by BMP-4 expression during neuronal differentiation in P19 cells[J]. Mol Cell Biochem, 2016, 412(1-2): 49-57.
[25] 姜明宇,任明永,刘海春,等. BMP4和拮抗剂Noggin在显性脊柱裂胎鼠发病中的作用[J].中国妇幼保健杂志,2017,32(20):5122-5123.
[26] Janowska J, Gargas J, Ziemka-Nalecz M, et al. Directed glial differentiation and transdifferentiation for neural tissue regeneration[J]. Exp Neurol, 2018, 319: 112813. doi: 10.1016/j.expneurol.2018.08.010.
[1] 杨晓倩 季静 刘娜 郭冬梅 崔癉. 三氯化铁及络合铁的抗银屑病作用研究[J]. 山东大学学报(医学版), 2209, 47(6): 114-117.
[2] 段淑红 刘凯 尹海燕 赵世斗 刘丰韬. 凝集素受体WGA、RCA和ECL在过量维甲酸致昆明小鼠腭裂发生中的作用[J]. 山东大学学报(医学版), 2209, 47(6): 47-.
[3] 张秀芳,李沛铮,张博涵,孙丛丛,刘艺鸣. 生长分化因子15在LPS诱导的帕金森病模型中的保护作用及机制[J]. 山东大学学报 (医学版), 2022, 60(5): 1-7.
[4] 刘敏,张玉超,马小莉,刘昕宇,孙露,左丹,刘元涛. 孤核受体NR4A1在H2O2诱导小鼠肾脏足细胞损伤中的作用[J]. 山东大学学报 (医学版), 2022, 60(5): 16-21.
[5] 菅天孜,陈诺,李理想,李延青,李艳. D-甘露糖和葡萄糖在溃疡性结肠炎小鼠中的作用[J]. 山东大学学报 (医学版), 2022, 60(3): 24-28.
[6] 隋荣翠,韩书慧,张宪昭,范新泰,王娜,侯凌霄,许安廷. 小鼠内淋巴囊原代上皮细胞L型钙离子通道定位表达[J]. 山东大学学报 (医学版), 2021, 59(10): 17-22.
[7] 索东阳,申飞,郭皓,刘力畅,杨惠敏,杨向东. Tim-3在药物性急性肾损伤动物模型中的表达及作用机制[J]. 山东大学学报 (医学版), 2020, 1(7): 1-6.
[8] 苏晓慧,孙明琪,张梅洁,方丽,罗丹丹,陈国平,于春晓,管庆波. 穗加精液分析自动检测系统与血细胞计数板人工计数法在小鼠精液分析中的应用[J]. 山东大学学报 (医学版), 2018, 56(6): 1-5.
[9] 焦凤萍,王玉,于爱莲,田兆菊,杨树林. 重组质粒pcDNA3.1(-)-P6-gBCTL-TBK-1的免疫效果评价[J]. 山东大学学报（医学版）, 2017, 55(3): 75-78.
[10] 李洪志,申永超,刘洁婷,赵孝金,初彦辉,袁晓环. 11β-HSD1抑制剂改善db/db小鼠胰岛素敏感性的实验研究[J]. 山东大学学报（医学版）, 2017, 55(10): 59-64.
[11] 郑倩倩,刘倜,谢克勤,林艺,李忠,王显军,毕振强,温红玲,宋艳艳,赵丽. 大蒜油在小鼠体内抗流感病毒作用[J]. 山东大学学报（医学版）, 2016, 54(7): 23-27.
[12] 赵栋燕,于岩波,左秀丽. 脑源性神经营养因子在小鼠肠黏膜屏障中的作用[J]. 山东大学学报（医学版）, 2016, 54(7): 1-5.
[13] 赵秀芹,邓春颖,李世英,张晋霞,贺永贵,余红,刘斌. 丁苯酞注射液对大鼠局灶性脑缺血再灌注损伤的神经保护作用[J]. 山东大学学报（医学版）, 2016, 54(4): 25-31.
[14] 李懋禹,唐诗,杜怡峰. TRB3在APP/PS1转基因小鼠脑内的表达及其意义[J]. 山东大学学报（医学版）, 2016, 54(10): 1-5.
[15] 贾俊伟,周雪颖,李秋红,于凯,周国钰,周盛年. Annexin A7在匹罗卡品癫痫小鼠模型脑组织中的表达[J]. 山东大学学报（医学版）, 2016, 54(10): 6-10.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 黄庆,田辉,李林,梁飞,刘贤锡 . 老年人肺癌组织中鸟氨酸脱羧酶基因表达及其临床意义[J]. 山东大学学报(医学版), 2006, 44(6): 556 -559 .
[2] 朱梅佳,韩巨,王新怡,鹿伟,王爱华,关心华,曹霞,曹秉振. 伴有皮层下梗死和白质脑病的常染色体显性遗传性脑动脉病临床病理研究[J]. 山东大学学报(医学版), 2006, 44(8): 834 -839 .
[3] 于慧1,2 ，陈少华1 ，赵家军2 ，高聆3
. 乙醇对人肝L02细胞糖原和GSK3β、PAMPK的影响[J]. 山东大学学报(医学版), 2009, 47(04): 75 -78 .
[4] 曾季平,王丽娜,王立祥,任晓辉,张孟业,夏文,崔行. 氯化锰致PC12细胞损伤的研究[J]. 山东大学学报(医学版), 2006, 44(5): 467 -470 .
[5] 舒雅,齐峰. 下颌角托在肥胖患者全麻诱导中的应用[J]. 山东大学学报(医学版), 2007, 45(10): 1072 -1074 .
[6] 宋海岩,武玉玲,张艳萍. 牡蛎提取物对高温致神经管畸形中凋亡细胞的保护作用[J]. 山东大学学报(医学版), 2007, 45(2): 113 -116 .
[7] 赵正斌1，薛双林2，张立婷1，李俊峰1，赵荣荣1，周海莲3，陈红1. 原花青素对瘦素诱导肝星状细胞增殖和TIMP-1产生的影响[J]. 山东大学学报(医学版), 2012, 50(7): 46 .
[8] 王志刚,丁　璇,孙　鹏/sup>,王成伟,郝晓光,潘　顺 . 术前脑血管造影在血管内支架成形术治疗缺血性脑血管病中的应用[J]. 山东大学学报(医学版), 2007, 45(2): 146 -148 .
[9] 孟静1，刘晓立2，郭春3，马春红3 . 小鼠Tim-３真核表达载体pTARGET-Tim-3的构建[J]. 山东大学学报(医学版), 2010, 48(11): 37 -40 .
[10] 郑金旭 顾燕兰 万兵. 糜酶抑制剂对肺纤维化的干预作用及其机制研究[J]. 山东大学学报(医学版), 2008, 46(12): 1132 -1135 .
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发