镇心省睡益智方及其精油对AD模型小鼠学习记忆的影响
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摘要
目的:观察镇心省睡益智方水提液及其精油对β淀粉样前体蛋白基因/早老素基因(APP/PS1)双转基因小鼠学习记忆的影响并探讨其可能机制。方法:采用APP/PS1双转基因痴呆小鼠及同月龄相同遗传背景C57BL/6JNju小鼠2种小鼠。C57BL/6JNju小鼠作为正常组,APP/PS1双转基因痴呆小鼠随机分为模型组,镇心省睡益智方精油低、高质量浓度组(12.13,48.50 mg·L~(-1)),镇心省睡益智方水提液组(0.46 g·kg~(-1)),每组12只。每天给药1次,连续给药22 d。给药结束后采用跳台实验、Morris水迷宫实验对小鼠行为学能力进行检测,采用尼氏染色观察海马CA1区神经元变化,采用硫磺素(Th S)染色观察海马DG区老年斑(SP)沉积,采用免疫组化法检测小鼠脑组织中葡萄糖转运蛋白1(GLUT1),胰岛素受体底物-1(IRS-1)的表达,采用酶联免疫吸附测定(ELISA)检测小鼠海马组织中乙酰胆碱(ACH),γ-氨基丁酸(GABA),谷氨酸(GLU)含量的变化。结果:与正常组比较,模型组小鼠跳台实验潜伏期显著缩短,错误次数显著增加(P<0.01),Morris水迷宫实验定位航行逃避潜伏期明显延长(P<0.05,P<0.01),海马CA1区神经元出现缺失,DG区出现明显的老年斑沉积(P<0.05),ACH,GLUT1含量均显著下降(P<0.01),GABA,GLU水平及IRS-1的表达均显著升高(P<0.01);与模型组比较,各给药组均可显著延长小鼠跳台实验潜伏期、减少跳台错误次数(P<0.01),明显降低定位航行小鼠逃避潜伏期(P<0.05,P<0.01),可一定程度保护小鼠海马CA1区神经元,减少DG区老年斑沉积(P<0.05,P<0.01),明显增加小鼠脑组织中ACH,GLUT1表达(P<0.05,P<0.01),显著降低GABA,GLU水平及IRS-1的表达(P<0.01)。结论:镇心省睡益智方水提液及其精油能够改善APP/PS1小鼠的学习记忆行为,保护神经元,增加脑组织GLUT1的表达,减少脑组织IRS-1的表达,减少老年斑沉积,升高ACH含量,降低GABA,GLU含量,可能是其防治阿尔茨海默症的机制。
Objective: The present study was designed to investigate the modulating effect of Zhenxin Xingshui Yizhi Fang and its essential oil extract on cognitive deficits in mice. Method: For the purpose of this study 5 months old APP/PS1 double transgenic mice and wild-type C57BL/6JNju were selected as experimental animals. Then APP/PS1 double transgenic mice were randomly divided into model group,essential oil low and high-dose groups( 12. 13,48. 50 mg·L~(-1)),Zhenxin Xingshui Yizhi Fang group( 0. 46 g·kg~(-1)). Meanwhile,wild-type C57BL/6JNju mice were used as a normal group. APP/PS1 double transgenic mice were treated with Zhenxin Xingshui Yizhi Fang and its essential oil extract for 22 consecutive days. Mice were subjected to a Morris water maze test and a platform test in order to determine their cognitive effect. Nissl’s staining was used to observe pathological changes in brain tissue. Meanwhile,senile plaques( SP) were observed by employing Thioflavin-S staining. The expression of glucose transporter 1( GLUT1) and insulin receptor substrate-1( IRS-1) were analyzed using immunohistochemistry techniques. The levels of neurotransmitters such as acetylcholine( ACH),glutamate( GLU) and γ-aminobutyric acid( GABA) in the hippocampus were quantified by enzyme-linked immunosorbent assay( ELISA). Result: The memory function was significantly reduced in model group,and severe brain injury and neuronal apoptosis were also observed in comparison to normal group( P < 0. 05,P < 0. 01). Compared with the model group,Zhenxin Xingshui Yizhi Fang and its essential oil extract improved previous learning and memory impairments( P < 0. 05,P < 0. 01),and the levels of SP,IRS-1,GLU,GABA( except essential oil low dose group) were significantly reduced( P < 0. 01),meanwhile the levels of ACH and GLUT1 were significantly improved( P < 0. 05,P < 0. 01). Conclusion: These results indicate that Zhenxin Xingshui Yizhi Fang and its essential oil extract could ameliorate cognitive deficits and GLUT1 and IRS-1 could be a possible therapeutic target for AD. It may be an interesting approach to the treatment of Alzheimer’s disease.
Objective: The present study was designed to investigate the modulating effect of Zhenxin Xingshui Yizhi Fang and its essential oil extract on cognitive deficits in mice. Method: For the purpose of this study 5 months old APP/PS1 double transgenic mice and wild-type C57BL/6JNju were selected as experimental animals. Then APP/PS1 double transgenic mice were randomly divided into model group,essential oil low and high-dose groups( 12. 13,48. 50 mg·L~(-1)),Zhenxin Xingshui Yizhi Fang group( 0. 46 g·kg~(-1)). Meanwhile,wild-type C57BL/6JNju mice were used as a normal group. APP/PS1 double transgenic mice were treated with Zhenxin Xingshui Yizhi Fang and its essential oil extract for 22 consecutive days. Mice were subjected to a Morris water maze test and a platform test in order to determine their cognitive effect. Nissl’s staining was used to observe pathological changes in brain tissue. Meanwhile,senile plaques( SP) were observed by employing Thioflavin-S staining. The expression of glucose transporter 1( GLUT1) and insulin receptor substrate-1( IRS-1) were analyzed using immunohistochemistry techniques. The levels of neurotransmitters such as acetylcholine( ACH),glutamate( GLU) and γ-aminobutyric acid( GABA) in the hippocampus were quantified by enzyme-linked immunosorbent assay( ELISA). Result: The memory function was significantly reduced in model group,and severe brain injury and neuronal apoptosis were also observed in comparison to normal group( P < 0. 05,P < 0. 01). Compared with the model group,Zhenxin Xingshui Yizhi Fang and its essential oil extract improved previous learning and memory impairments( P < 0. 05,P < 0. 01),and the levels of SP,IRS-1,GLU,GABA( except essential oil low dose group) were significantly reduced( P < 0. 01),meanwhile the levels of ACH and GLUT1 were significantly improved( P < 0. 05,P < 0. 01). Conclusion: These results indicate that Zhenxin Xingshui Yizhi Fang and its essential oil extract could ameliorate cognitive deficits and GLUT1 and IRS-1 could be a possible therapeutic target for AD. It may be an interesting approach to the treatment of Alzheimer’s disease.
引文
[1]PENG D,YUAN X,ZHU R.Memantine hydrochloride in the treatment of dementia subtypes[J].J Clin Neurosci,2013,20(11):1482-1485.
[2]张雅静,纪勇,石志鸿.阿尔茨海默病的药物治疗进展[J].中国城乡企业卫生,2016,31(5):15-18.
[3]毕祥云,陈学香.阿尔茨海默病相关治疗的研究进展[J].中华保健医学杂志,2017,19(3):275-277.
[4]Demetrius L A,Driver J.Alzheimer's as a metabolic disease[J].Biogerontology,2013,14(6):641-649.
[5]Monte S M D L,TONG M.Brain metabolic dysfunction at the core of Alzheimer's disease[J].Biochem Pharmacol,2014,88(4):548-559.
[6]Blonz E R.Alzheimer's disease as the product of a progressive energy deficiency syndrome in the central nervous system:the neuroenergetic hypothesis[J].Alzheimers Dis,2017,60(4):1223-1229.
[7]Duara R,Grady C,Haxby J,et al.Positron emission tomography in Alzheimer's disease[J].Neurology,1986,36(7):879-887.
[8]Denver P,English A,Mcclean P L.Inflammation,insulin signaling and cognitive function in aged APP/PS1 mice[J].Brain Behav Immun,2018,doi:10.1016/j.bbi.2018.03.032.
[9]Choeiri C,Staines W,Messier C.Immunohistochemical localization and quantification of glucose transporters in the mouse brain[J].Neurosci,2002,111(1):19-34.
[10][唐]孙思邈.千金翼方[M].北京:人民卫生出版社,1983:188.
[11]范昌斌,杨旭东.镇心醒睡益智方治疗嗜睡症14例[J].基层中药杂志,2000,14(3):61.
[12]贺密会,刘明,周小江,等.镇心省睡益智方对快动眼睡眠剥夺大鼠促醒和认知功能的影响[J].解放军药学学报,2013,29(5):415-419.
[13]杭天依,李光武,徐金勇.嗅觉通路与大脑关联作用的研究进展[J].立体定向和功能性神经外科杂志,2013,26(2):125-128.
[14]谭洁,罗敏敏.嗅球对嗅觉信息的处理[J].生物物理学报,2010,26(3):194-208.
[15]Dobetsberger C,Buchbauer G.Actions of essential oils on the central nervous system:an updated review[J].Flavour Fragr J,2011,26(5):300-316.
[16]Moss L,Rouse M,Wesnes K A,et al.Differential effects of the aromas of salvia species on memory and mood[J].Hum Psychopharmacol Clin Exp,2010,25(5):388-396.
[17]Jimbo D,Kimura Y,Taniguchi M,et al.Effect of aromatherapy on patients with Alzheimer's disease[J].Psychogeriatrics,2010,9(4):173-179.
[18]胡泓博,夏传余,李光武.丁香酚吸嗅对MCAO模型大鼠脑源性神经营养因子的影响[J].中风与神经疾病杂志,2015,32(1):28-32.
[19]徐叔云,卞如濂,陈修.《药理实验方法学》第二版出版发行[J].中国药理学通报,1992(1):19.
[20]魏伟,吴希美,李元建.药理实验方法学[M].北京:人民卫生出版社,2010:19.
[21]闫盼盼,闫国立,詹向红,等.Morris水迷宫实验设计的统计学方法探析[J].中华中医药学刊,2013,31(2):264-266.
[22]Kalita J,Kumar V,Misra U K,et al.Memory and learning dysfunction following copper toxicity:biochemical and immunohistochemical basis[J].Mol Neurobiol,2017,55(2):1-12.
[23]Kim H Y,Kim H V,Jo S,et al.EPPS rescues hippocampus-dependent cognitive deficits in APP/PS1mice by disaggregation of amyloid-βoligomers and plaques[J].Nat Commun,2016,7(8997):10755.
[24]张玲.HDAC6在APPswe/PS1d E9阿尔茨海默病转基因模型小鼠发病机制中的作用[D].北京:北京协和医学院,2014.
[25]郝徐艺,罗思,程淑意,等.当归芍药散对AD细胞模型铜离子介导的Aβ聚集的影响[J].中国实验方剂学杂志,2019,25(6):45-51.
[26]Minkeviciene R,Ihalainen J,Malm T,et al.Agerelated decrease in stimulated glutamate release and vesicular glutamate transporters in APP/PS1 transgenic and wild-type mice[J].J Neurochem,2008,105(3):584-594.
[27]Hashimoto E,Ozawa H,Saito T,et al.Impairment of Gsαfunction in human brain cortex of Alzheimer's disease:comparison with normal aging[J].J Neural Transm,2004,111(3):311-322.
[28]Patching S G.Glucose transporters at the blood-brain barrier:function,regulation and gateways for drug delivery[J].Mol Neurobiol,2016,54(2):1046-1077.
[29]Szutowicz A,Bielarczyk H,Ronowska A,et al.Intracellular redistribution of acetyl-Co A,the pivotal point in differential susceptibility of cholinergic neurons and glial cells to neurodegenerative signals[J].Biochem Soc T,2014,42(4):1101-1106.
[30]PAN J,KAO Y L,Joshi S,et al.Activation of Rac1 by phosphatidylinositol 3-kinase in vivo:role in activation of mitogen-activated protein kinase(MAPK)pathways and retinoic acid-induced neuronal differentiation of SHSY5Y cells[J].J Neurochem,2010,93(3):571-583.
[31]何瑛琨.基于脑能量代谢研究半夏泻心汤的神经保护作用[D].北京:北京中医药大学,2016.
[32]卢成淑,冯宁,南国,等.石菖蒲及其活性成分防治阿尔茨海默病的研究进展[J].中草药,2016,47(7):1236-1242.
[33]GENG Y,LI C,LIU J,et al.Beta-asarone improves cognitive function by suppressing neuronal apoptosis in the beta-amyloid hippocampus injection rats[J].Biol Pharm Bull,2010,33(5):836-843.
[34]金虹,徐国波,黄毅.石菖蒲抑酶活性的实验研究[J].时珍国医国药,2009,20(9):2241-2242.
[35]吴宾,方永奇.石菖蒲益智作用的物质基础及其机理研究[J].中医药学刊,2004,22(9):1635-1636,1641.
[36]杨天鹏,唐敏,刘巧琼,等.丁香酚通过嗅觉通路改善昆明鼠学习记忆的机制[J].中国康复医学杂志,2007,22(6):487-489,508,477.
[37]张陶珍,荣巍巍,李清,等.远志的研究进展[J].中草药,2016,47(13):2381-2389.
[2]张雅静,纪勇,石志鸿.阿尔茨海默病的药物治疗进展[J].中国城乡企业卫生,2016,31(5):15-18.
[3]毕祥云,陈学香.阿尔茨海默病相关治疗的研究进展[J].中华保健医学杂志,2017,19(3):275-277.
[4]Demetrius L A,Driver J.Alzheimer's as a metabolic disease[J].Biogerontology,2013,14(6):641-649.
[5]Monte S M D L,TONG M.Brain metabolic dysfunction at the core of Alzheimer's disease[J].Biochem Pharmacol,2014,88(4):548-559.
[6]Blonz E R.Alzheimer's disease as the product of a progressive energy deficiency syndrome in the central nervous system:the neuroenergetic hypothesis[J].Alzheimers Dis,2017,60(4):1223-1229.
[7]Duara R,Grady C,Haxby J,et al.Positron emission tomography in Alzheimer's disease[J].Neurology,1986,36(7):879-887.
[8]Denver P,English A,Mcclean P L.Inflammation,insulin signaling and cognitive function in aged APP/PS1 mice[J].Brain Behav Immun,2018,doi:10.1016/j.bbi.2018.03.032.
[9]Choeiri C,Staines W,Messier C.Immunohistochemical localization and quantification of glucose transporters in the mouse brain[J].Neurosci,2002,111(1):19-34.
[10][唐]孙思邈.千金翼方[M].北京:人民卫生出版社,1983:188.
[11]范昌斌,杨旭东.镇心醒睡益智方治疗嗜睡症14例[J].基层中药杂志,2000,14(3):61.
[12]贺密会,刘明,周小江,等.镇心省睡益智方对快动眼睡眠剥夺大鼠促醒和认知功能的影响[J].解放军药学学报,2013,29(5):415-419.
[13]杭天依,李光武,徐金勇.嗅觉通路与大脑关联作用的研究进展[J].立体定向和功能性神经外科杂志,2013,26(2):125-128.
[14]谭洁,罗敏敏.嗅球对嗅觉信息的处理[J].生物物理学报,2010,26(3):194-208.
[15]Dobetsberger C,Buchbauer G.Actions of essential oils on the central nervous system:an updated review[J].Flavour Fragr J,2011,26(5):300-316.
[16]Moss L,Rouse M,Wesnes K A,et al.Differential effects of the aromas of salvia species on memory and mood[J].Hum Psychopharmacol Clin Exp,2010,25(5):388-396.
[17]Jimbo D,Kimura Y,Taniguchi M,et al.Effect of aromatherapy on patients with Alzheimer's disease[J].Psychogeriatrics,2010,9(4):173-179.
[18]胡泓博,夏传余,李光武.丁香酚吸嗅对MCAO模型大鼠脑源性神经营养因子的影响[J].中风与神经疾病杂志,2015,32(1):28-32.
[19]徐叔云,卞如濂,陈修.《药理实验方法学》第二版出版发行[J].中国药理学通报,1992(1):19.
[20]魏伟,吴希美,李元建.药理实验方法学[M].北京:人民卫生出版社,2010:19.
[21]闫盼盼,闫国立,詹向红,等.Morris水迷宫实验设计的统计学方法探析[J].中华中医药学刊,2013,31(2):264-266.
[22]Kalita J,Kumar V,Misra U K,et al.Memory and learning dysfunction following copper toxicity:biochemical and immunohistochemical basis[J].Mol Neurobiol,2017,55(2):1-12.
[23]Kim H Y,Kim H V,Jo S,et al.EPPS rescues hippocampus-dependent cognitive deficits in APP/PS1mice by disaggregation of amyloid-βoligomers and plaques[J].Nat Commun,2016,7(8997):10755.
[24]张玲.HDAC6在APPswe/PS1d E9阿尔茨海默病转基因模型小鼠发病机制中的作用[D].北京:北京协和医学院,2014.
[25]郝徐艺,罗思,程淑意,等.当归芍药散对AD细胞模型铜离子介导的Aβ聚集的影响[J].中国实验方剂学杂志,2019,25(6):45-51.
[26]Minkeviciene R,Ihalainen J,Malm T,et al.Agerelated decrease in stimulated glutamate release and vesicular glutamate transporters in APP/PS1 transgenic and wild-type mice[J].J Neurochem,2008,105(3):584-594.
[27]Hashimoto E,Ozawa H,Saito T,et al.Impairment of Gsαfunction in human brain cortex of Alzheimer's disease:comparison with normal aging[J].J Neural Transm,2004,111(3):311-322.
[28]Patching S G.Glucose transporters at the blood-brain barrier:function,regulation and gateways for drug delivery[J].Mol Neurobiol,2016,54(2):1046-1077.
[29]Szutowicz A,Bielarczyk H,Ronowska A,et al.Intracellular redistribution of acetyl-Co A,the pivotal point in differential susceptibility of cholinergic neurons and glial cells to neurodegenerative signals[J].Biochem Soc T,2014,42(4):1101-1106.
[30]PAN J,KAO Y L,Joshi S,et al.Activation of Rac1 by phosphatidylinositol 3-kinase in vivo:role in activation of mitogen-activated protein kinase(MAPK)pathways and retinoic acid-induced neuronal differentiation of SHSY5Y cells[J].J Neurochem,2010,93(3):571-583.
[31]何瑛琨.基于脑能量代谢研究半夏泻心汤的神经保护作用[D].北京:北京中医药大学,2016.
[32]卢成淑,冯宁,南国,等.石菖蒲及其活性成分防治阿尔茨海默病的研究进展[J].中草药,2016,47(7):1236-1242.
[33]GENG Y,LI C,LIU J,et al.Beta-asarone improves cognitive function by suppressing neuronal apoptosis in the beta-amyloid hippocampus injection rats[J].Biol Pharm Bull,2010,33(5):836-843.
[34]金虹,徐国波,黄毅.石菖蒲抑酶活性的实验研究[J].时珍国医国药,2009,20(9):2241-2242.
[35]吴宾,方永奇.石菖蒲益智作用的物质基础及其机理研究[J].中医药学刊,2004,22(9):1635-1636,1641.
[36]杨天鹏,唐敏,刘巧琼,等.丁香酚通过嗅觉通路改善昆明鼠学习记忆的机制[J].中国康复医学杂志,2007,22(6):487-489,508,477.
[37]张陶珍,荣巍巍,李清,等.远志的研究进展[J].中草药,2016,47(13):2381-2389.