Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2018, Vol. 47 Issue (5): 450-456    DOI: 10.3785/j.issn.1008-9292.2018.10.02
专题报道     
和厚朴酚对癫痫小鼠学习记忆能力的改善作用
王青梅1(),舒敏1,徐千姿1,谢一乙1,阮盛哲1,王健达2,*(),曾玲晖1,*()
1. 浙江大学城市学院医学院, 浙江 杭州 310015
2. 浙江大学医学院附属第二医院儿科, 浙江 杭州 310012
Effects of Honokiol on cognitive function in mice with kainic acid-induced epilepsy
WANG Qingmei1(),SHU Min1,XU Qianzi1,XIE Yiyi1,RUAN Shengzhe1,WANG Jianda2,*(),ZENG Linghui1,*()
1. School of Medicine, Zhejiang University City College, Hangzhou 310015, China
2. Department of Pediatrics, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China
 全文: PDF(1342 KB)   HTML( 14 )
摘要：

目的: 探索和厚朴酚对癫痫小鼠学习记忆能力的改善作用并探讨其机制。方法: 5周龄雄性ICR小鼠采用腹腔注射红藻氨酸（38 mg/kg）的方式建立癫痫模型。治疗组小鼠腹腔注射和厚朴酚（3、10、30 mg/kg）10 d。Fluoro-Jade B染色法检测神经元活性；Morris水迷宫和Y迷宫实验观察小鼠学习记忆能力；蛋白质印迹法检测乙酰化超氧化物歧化酶（SOD）、微管相关蛋白1轻链3-Ⅱ（LC3-Ⅱ）和P62蛋白表达水平；硫代巴比妥酸法和WST-1法分别测定丙二醛和SOD等氧化应激产物的含量。结果: 与对照组比较，模型组神经元凋亡数量增加，学习记忆能力下降，乙酰化SOD和丙二醛表达量增加，SOD活性下降，LC3-Ⅱ和P62蛋白表达量增加（P < 0.05或P < 0.01）；与模型组比较，和厚朴酚10 mg/kg和30 mg/kg均可减少神经元凋亡数量，改善学习记忆能力，并可逆转癫痫发作所致的丙二醛和SOD乙酰化水平，降低LC3-Ⅱ和P62的表达量（P < 0.05或P < 0.01），且药物剂量为30 mg/kg时效果更加显著。结论: 和厚朴酚可缓解癫痫发作导致的氧化应激和自噬降解障碍，减少神经元凋亡，从而改善癫痫小鼠的学习记忆能力。
关键词： 和厚朴酚/药理学;  癫痫/药物疗法;  记忆/药物作用;  学习/药物作用;  氧化应激;  自噬;  小鼠, 近交ICR    
Abstract:

Objective: To investigate the effects of Honokiol on cognitive function in mice with epilepsy. Methods: Kainic acid (38 mg/kg) was intraperitoneally injected in 5 weeks old male ICR mice to induce epilepsy. Honokiol at dose of 3, 10, 30 mg/kg was given to epilepic mice by intraperitoneal injection for 10 days. Fluoro-Jade B staining was used to assess neuronal death; Morris water maze and Y maze tests were used to measure cognitive function such as learning and memory; Western blot was performed to detect the expression of acetylated superoxide dismutase (SOD), microtubule associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ) and P62 in hippocampus tissue; thiobarbituric acid and WST-1 methods were used to detect malondialdehyde (MDA) and SOD. Results: Compared with control group, the levels of acetylated-SOD, MDA, LC3-Ⅱ, P62 and neuronal death increased, cognitive function and SOD decreased in model group (P < 0.05 or P < 0.01). Honokiol at the dose of 10 mg/kg and 30 mg/kg decreased SOD acetylation, MDA content, expression of LC3-Ⅱ and P62, as well as neuronal death, and the cognitive function was improved (P < 0.05 or P < 0.01), especially in 30 mg/kg Honokiol group. Conclusion: Honokiol alleviates oxidative stress and autophagy degradation disorder, decreases neuronal death, and therefore improves cognitive function in epilepsy mice.
Key words: Honokiol/pharmacology    Epilepsy/drug therapy    Memory/drug effects    Learning/drug effects    Oxidative stress    Autophagy    Mice, inbred ICR
收稿日期: 2018-09-12 出版日期: 2019-01-23
:  R742.1  
基金资助: 杭州市科技发展计划重大科技创新专项(20152013A02);国家自然科学基金(81600988)
通讯作者: 王健达,曾玲晖     E-mail: 2747912115@qq.com;2515165@zju.edu.cn;zenglh@zucc.edu.cn
作者简介: 王青梅(1994-), 女, 硕士研究生, 主要从事药理学研究; E-mail:2747912115@qq.com; https://orcid.org/0000-0002-3220-6762
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
王青梅
舒敏
徐千姿
谢一乙
阮盛哲
王健达
曾玲晖

引用本文:

王青梅,舒敏,徐千姿,谢一乙,阮盛哲,王健达,曾玲晖. 和厚朴酚对癫痫小鼠学习记忆能力的改善作用[J]. 浙江大学学报（医学版）, 2018, 47(5): 450-456.

WANG Qingmei,SHU Min,XU Qianzi,XIE Yiyi,RUAN Shengzhe,WANG Jianda,ZENG Linghui. Effects of Honokiol on cognitive function in mice with kainic acid-induced epilepsy. J Zhejiang Univ (Med Sci), 2018, 47(5): 450-456.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2018.10.02    或    http://www.zjujournals.com/med/CN/Y2018/V47/I5/450

图 1  各组海马组织中神经元Fluoro-Jade B(FJB)染色荧光图
图 2  各组Morris水迷宫实验结果比较(n=10)
图 3  各组Y迷宫实验结果比较(n=10)
图 4  各组海马组织中乙酰化SOD表达电泳图
图 5  各组海马组织中氧化应激产物表达量比较(n=6)
图 6  各组海马组织中LC3-Ⅱ和P62蛋白表达及比较(n=10)
1 NGUGI A K , BOTTOMLEY C , FEGAN G et al. Premature mortality in active convulsive epilepsy in rural Kenya:causes and associated factors[J]. Neurology, 2014, 82 (7): 582- 589
doi: 10.1212/WNL.0000000000000123
2 HOLMES G L . Effects of early seizures on later behavior and epileptogenicity[J]. Ment Retard Dev Disabil Res Rev, 2004, 10 (2): 101- 105
doi: 10.1002/(ISSN)1098-2779
3 LEE S E , KIBBY M Y , COHEN M J et al. Differences in memory functioning between children with attention-deficit/hyperactivity disorder and/or focal epilepsy[J]. Child Neuropsychol, 2016, 22 (8): 979- 1000
doi: 10.1080/09297049.2015.1060955
4 SKIRROW C , CROSS J H , HARRISON S et al. Temporal lobe surgery in childhood and neuroanatomical predictors of long-term declarative memory outcome[J]. Brain, 2015, 138 (Pt 1): 80- 93
5 LIN J J , MULA M , HERMANN B P . Uncovering the neurobehavioral comorbidities of epilepsy over the lifespan[J]. Lancet, 2012, 380 (9848): 1180- 1192
doi: 10.1016/S0140-6736(12)61455-X
6 COYLE J T , PUTTFARCKEN P . Oxidative stress, glutamate, and neurodegenerative disorders[J]. Science, 1993, 262 (5134): 689- 695
doi: 10.1126/science.7901908
7 FRANTSEVA M V , VELAZQUEZ J L , HWANG P A et al. Free radical production correlates with cell death in an in vitro model of epilepsy[J]. Eur J Neurosci, 2000, 12 (4): 1431- 1439
doi: 10.1046/j.1460-9568.2000.00016.x
8 CHEN W , SUN Y , LIU K et al. Autophagy:a double-edged sword for neuronal survival after cerebral ischemia[J]. Neural Regen Res, 2014, 9 (12): 1210- 1216
doi: 10.4103/1673-5374.135329
9 NILSSON P , SAIDO T C . Dual roles for autophagy:Degradation and secretion of Alzheimer's disease Aβ peptide[J]. Bioessays, 2014, 36 (6): 570- 578
doi: 10.1002/bies.201400002
10 MCMAHON J , HUANG X , YANG J et al. Impaired autophagy in neurons after disinhibition of mammalian target of rapamycin and its contribution to epileptogenesis[J]. J Neurosci, 2012, 32 (45): 15704- 15714
doi: 10.1523/JNEUROSCI.2392-12.2012
11 郝庆红, 陈冠华, 冯雅琪 et al. 厚朴中清除氧自由基主要有效成分的确定[J]. 河北大学学报(自然科学版), 2005, 25 (2): 164- 166, 178
HAO Qinghong , CHEN Guanhua , FENG Yaqi et al. Determination of the main efficient components in the herb of magnolia officinalis for eliminating oxygen radicals[J]. Journal of Hebei University(Natural Science Edition), 2005, 25 (2): 164- 166, 178
doi: 10.3969/j.issn.1000-1565.2005.02.011
12 HAHM E R , SAKAO K , SINGH S V . Honokiol activates reactive oxygen species-mediated cytoprotective autophagy in human prostate cancer cells[J]. Prostate, 2014, 74 (12): 1209- 1221
doi: 10.1002/pros.v74.12
13 RACINE R J . Modification of seizure activity by electrical stimulation. Ⅱ. Motor seizure[J]. Electroencephalogr Clin Neurophysiol, 1972, 32 (3): 281- 294
doi: 10.1016/0013-4694(72)90177-0
14 ZENG L H , RENSING N R , WONG M . The mammalian target of rapamycin signaling pathway mediates epileptogenesis in a model of temporal lobe epilepsy[J]. J Neurosci, 2009, 29 (21): 6964- 6972
doi: 10.1523/JNEUROSCI.0066-09.2009
15 SHARMA S , RAKOCZY S , BROWN-BORG H . Assessment of spatial memory in mice[J]. Life Sci, 2010, 87 (17-18): 521- 536
doi: 10.1016/j.lfs.2010.09.004
16 HOLLEY A J , LUGO J N . Effects of an acute seizure on associative learning and memory[J]. Epilepsy Behav, 2016, 54:51- 57
doi: 10.1016/j.yebeh.2015.11.001
17 BOWER M R , STEAD M , BOWER R S. et al. Evidence for consolidation of neuronal assemblies after seizures in humans[J]. J Neurosci, 2015, 35 (3): 999- 1010
doi: 10.1523/JNEUROSCI.3019-14.2015
18 KOVAC S, DINKOVA KOSTOVA A T, HERRMANN A M, et al. Metabolic and homeostatic changes in seizures and acquired epilepsy-mitochondria, calcium dynamics and reactive oxygen species[J/OL]. Int J Mol Sci, 2017, 18(9): E1935.
19 WOJSIAG J , ZOLTOWSKA K M , LASKOWSKA-KASZUB K et al. Oxidant/antioxidant imbalance in Alzheimer's disease:therapeutic and diagnostic prospects[J]. Oxid Med Cell Longev, 2018, 6435861
20 JIN Y N, HWANG W Y, JO C, et al. Metabolic state determines sensitivity to cellular stress in Huntington disease: normalization by activation of PPARγ[J/OL]. PLoS One, 2012, 7(1): e30406.
21 CHENG A , YANG Y , ZHOU Y et al. Mitochondrial SIRT3 mediates adaptive responses of neurons to exercise and metabolic and excitatory challenges[J]. Cell Metab, 2016, 23 (1): 128- 142
[1] 吴忧,梁顺利,徐彬,张荣博,徐林胜. 姜黄素保护帕金森病多巴胺能神经元的机制研究[J]. 浙江大学学报（医学版）, 2018, 47(5): 480-486.
[2] 朱锋,范苗,徐孜惟,蔡依廷,陈益臻,余双,曾玲晖. 雷帕霉素对帕金森病小鼠的保护作用[J]. 浙江大学学报（医学版）, 2018, 47(5): 465-472.
[3] 吕丹丹,应可净. 自噬在肺动脉高压发生和发展中的调节作用[J]. 浙江大学学报（医学版）, 2018, 47(2): 207-212.
[4] 张斌斌 等. 抑制哺乳动物雷帕霉素靶蛋白信号通路对慢性脑缺血小鼠认知功能的改善和机制[J]. 浙江大学学报（医学版）, 2017, 46(4): 405-412.
[5] 郑艳榕,张翔南,陈忠. Nix介导的线粒体自噬机制的研究进展[J]. 浙江大学学报（医学版）, 2017, 46(1): 92-96.
[6] 刘巧云 等. 锌与自噬[J]. 浙江大学学报（医学版）, 2016, 45(3): 308-314.
[7] 曹建平, 夏大静. 自噬与肿瘤关系研究新进展[J]. 浙江大学学报（医学版）, 2015, 44(2): 204-210.
[8] 刘健翔, 刘福荣, 杨鑫杰, 王昱植, 陈丹娇, 谢强敏, 曾玲晖. 甘草黄酮对红藻氨酸致痫小鼠的抗癫痫作用研究[J]. 浙江大学学报（医学版）, 2015, 44(1): 37-42.
[9] 张华丹，谢雅聪，翁灵，张宇辰，史琼瑶，陈涛，曾玲晖. 雷帕霉素对匹罗卡品诱导的癫痫小鼠学习记忆的改善作用[J]. 浙江大学学报（医学版）, 2013, 42(6): 602-608.
[10] 曾玲晖，丁美萍. mTOR信号通路在癫痫发生中的作用[J]. 浙江大学学报（医学版）, 2013, 42(6): 597-601.
[11] . 慢性铅暴露大鼠海马脑区自噬相关蛋白表达的研究[J]. 浙江大学学报（医学版）, 2012, 41(4): 402-409.
[12] . 紫心甘薯对高脂血症大鼠脂质代谢及氧化应激的影响[J]. 浙江大学学报（医学版）, 2011, 40(4): 360-364.
[13] 夏强，钱令波. 心脑缺血再灌注损伤的机制及防治策略研究进展[J]. 浙江大学学报（医学版）, 2010, 39(6): 551-558.
[14] 洪远,陈盛,张建民. 氢气对神经系统疾病治疗作用的研究进展[J]. 浙江大学学报（医学版）, 2010, 39(6): 638-643.
[15] 童茵，李璐，刘辉，钱文斌. 细胞自噬与血液肿瘤关系研究进展[J]. 浙江大学学报（医学版）, 2010, 39(3): 246-249.