电针对SAMP8小鼠血脑屏障的影响
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摘要
目的研究电针通过对快速老化痴呆模型SAMP8小鼠血脑屏障的调控,以期加强药物入脑。方法 30周龄雄性SAMP8小鼠随机分为模型组(n=7)、电针组(n=7)、药物组(n=7)和针药组(n=7),以同源抗快速老化小鼠SAMR1作为对照组(n=7)。电针组和针药组电针印堂、百会;药物组和针药组予盐酸多奈哌齐灌胃;对照组和模型组常规抓取。共4周。电镜观察血脑屏障紧密连接开放程度和基底膜厚度;定量聚合酶链式反应检测海马区紧密连接蛋白ZO-1、Claudin-5和Occludin mRNA的表达,免疫组化检测海马区乙酰胆碱酯酶(AChE)的含量。结果对照组、电针组和针药组紧密连接以及基底膜结构均优于模型组和药物组;针药组AChE表达最低,以下依次是药物组和对照组、电针组、模型组(P <0.001);电针组和针药组ZO-1、Claudin-5和Occludin mRNA表达水平高于药物组(P <0.01)。结论电针能够改善SAMP8小鼠血脑屏障结构,可能与调节紧密连接蛋白ZO-1、Claudin-5和Occludin的表达有关。
Objective To investigate the effects of electroacupuncture on blood-brain barrier to promote intake of drugs into brain of SAMP8 mice.Methods The male SAMP8 mice(30-week-old) were randomly divided into model group(n = 7), drug group(n = 7),acupuncture group(n = 7) and combined group(n = 7). Other SAMR1 mice were as control group(n = 7). The acupuncture group and the combined group accepted electroacupuncture at Baihui(DU20) and Yintang(EXHN3), the drug group and the combined group accepted Donepezil, for four weeks. They were observed hippo-campus tight junction(TJ) under transmission electron microscope. The activities of acetylcholinesterase(AChE)in hippocampus were detected with immunohistochemistry. The mRNA expression of ZO-1, Claudin-5 and Oc-cludin was detected with real-time quantitative polymerase chain reaction.Results TJ and basement membrane in the control group, the acupuncture group and the combined group were better than those in the model group and the drug group. AChE was the most in the combined group, and then the drug group and the control group, the acupuncture group and the model group(P < 0.001). The expression of mRNA of ZO-1, Claudin-5 and Occludin was more in the acupuncture group and the combined group than in the drug group(P < 0.01).Conclusion Electroacupuncture could ameliorate blood-brain barrier disruption and promote drug to enter the brain in SAMP8 mice, which may relate to the adjustion of ZO-1, Claudin-5 and Occludin.
Objective To investigate the effects of electroacupuncture on blood-brain barrier to promote intake of drugs into brain of SAMP8 mice.Methods The male SAMP8 mice(30-week-old) were randomly divided into model group(n = 7), drug group(n = 7),acupuncture group(n = 7) and combined group(n = 7). Other SAMR1 mice were as control group(n = 7). The acupuncture group and the combined group accepted electroacupuncture at Baihui(DU20) and Yintang(EXHN3), the drug group and the combined group accepted Donepezil, for four weeks. They were observed hippo-campus tight junction(TJ) under transmission electron microscope. The activities of acetylcholinesterase(AChE)in hippocampus were detected with immunohistochemistry. The mRNA expression of ZO-1, Claudin-5 and Oc-cludin was detected with real-time quantitative polymerase chain reaction.Results TJ and basement membrane in the control group, the acupuncture group and the combined group were better than those in the model group and the drug group. AChE was the most in the combined group, and then the drug group and the control group, the acupuncture group and the model group(P < 0.001). The expression of mRNA of ZO-1, Claudin-5 and Occludin was more in the acupuncture group and the combined group than in the drug group(P < 0.01).Conclusion Electroacupuncture could ameliorate blood-brain barrier disruption and promote drug to enter the brain in SAMP8 mice, which may relate to the adjustion of ZO-1, Claudin-5 and Occludin.
引文
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[5]李小宏,余和平,曹万群,等.电针联合安理申治疗轻度认知障碍的临床研究[J].激光杂志,2013,34(5):75-76.
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[8]王雪岩,周震,张琳琳,等.淫羊藿苷对SAMP8小鼠记忆能力、神经元微观形态及GSK-3β蛋白表达的影响[J].辽宁中医杂志,2014,41(12):2707-2710.
[9]Wu G,Li L,Li H,et al.Electroacupuncture ameliorates spatial learning and memory impairment via attenuating NOX2-related oxidative stress in a rat model of Alzheimer's disease induced by Aβ1-42[J].Mol Cell Biol,2017,63(4):38.
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[12]Zhang X,Li G,Guo L,et al.Age-related alteration in cerebral blood flow and energy failure is correlated with cognitive impairment in the senescence-accelerated prone mouse strain 8(SAMP8)[J].Neurol Sci,2013,34(11):1917-1924.
[13]Xiong H,Callaghan D,Jones A,et al.ABCG2 is upregulated in Al-zheimer's brain with cerebral amyloid angiopathy and may act as a gate-keeper at the blood-brain barrier for Aβ1-40 peptides[J].J Neurosci,2009,29(17):5463-5475.
[14]Di Marco L Y,Venneri A,Farkas E,et al.Vascular dysfunction in the pathogenesis of Alzheimer's disease-a review of endothelium-mediat-ed mechanisms and ensuing vicious circles[J].Neurobiol Dis,2015,82:593-606.
[15]Zlokovic B V.The blood-brain barrier in health and chronic neurode-generative disorders[J].Neuron,2008,57(2):178-201.
[16]Butterfield D A,Poon H F.The senescence-accelerated prone mouse(SAMP8):a model of age-related cognitive decline with relevance to al-terations of the gene expression and protein abnormalities in Alzheim-er's disease[J].Exp Gerontol,2005,40(10):774-783.
[17]Del Valle J,Duran-Vilaregut J,Manich G,et al.Cerebral amyloid an-giopathy,blood-brain barrier disruption and amyloid accumulation in SAMP8 mice[J].Neurodegener Dis,2011,8(6):421-429.
[18]杨清华,郭玲,陈清,等.电针刺激对APP/PS1双转基因阿尔兹海默病模型小鼠皮质老年斑形成的影响及机制研究[J].中国康复医学杂志,2018,33(7):770-776.
[19]Ning D,Jing J,Menghan L,et al.Manual acupuncture suppresses the expression of proinflammatory proteins associated with the NLRP3inflammasome in the hippocampus of SAMP8 mice[J].Evid Based Compl Alt,2017,2017:1-8.
[20]Reinhold A,Rittner H.Barrier function in the peripheral and central nervous system-a review[J].Eur J Appl Physiol,2016,469(1):1-12.
[21]廖华宁,汪宁,王艳.紧密连接蛋白与血脑屏障[J].安徽医药,2012,16(3):377-379.
[22]吴丽文,尹飞,彭镜,等.紧密连接蛋白ZO-1,occludin和actin参与缺氧缺血诱导的血脑屏障通透性增加[J].中国当代儿科杂志,2008,10(4):513-516.
[23]Gonzalez-Velasquez F J,Kotarek J A,Moss M A.Soluble aggregates of the amyloid-beta protein selectively stimulate permeability in human brain microvascular endothelial monolayers[J].J Neurochem,2008,107(2):466-477.
[24]Tai L M,Holloway K A,Male D K,et al.Amyloid-beta-induced oc-cludin down-regulation and increased permeability in human brain en-dothelial cells is mediated by MAPK activation[J].Cell Mol Med,2010,14(5):1101-1112.
[25]Gustafsson S,Gustavsson T,Roshanbin S,et al.Blood-brain barrier integrity in a mouse model of Alzheimer's disease with or without acute3D6 immunotherapy[J].Neuropharmacology,2018,143:1-9.
[26]Greta H,Stina S,Fang X T,et al.Bivalent brain shuttle increases an-tibody uptake by monovalent binding to the transferrin receptor[J].Theranostics,2017,7(2):308-318.
[27]Jung Y S,Lee S W,Park J H,et al.Electroacupuncture precondition-ing reduces ROS generation with NOX4 down-regulation and amelio-rates blood-brain barrier disruption after ischemic stroke[J].Biomed Sci,2016,23(1):32.
[28]孔海龙,韦旭,周倩,等.针药结合对阿尔茨海默病模型大鼠海马CA1区ChAT、AChE表达的影响[J].皖南医学院学报,2018,37(4):310-313.
[2]Mehta D C,Short J L,Hilmer S N,et al.Drug access to the central ner-vous system in Alzheimer's disease:preclinical and clinical insights[J].Pharm Res,2015,32(3):819-839.
[3]Mehta D C,Short J L,Nicolazzo J A.Altered brain uptake of therapeu-tics in a triple transgenic mouse model of Alzheimer's disease[J].Pharm Res,2013,30(11):2868-2879.
[4]Zipser B D,Johanson C E,Gonzalez L,et al.Microvascular injury and blood-brain barrier leakage in Alzheimer's disease[J].Neurobiol Ag-ing,2007,28(7):977-986.
[5]李小宏,余和平,曹万群,等.电针联合安理申治疗轻度认知障碍的临床研究[J].激光杂志,2013,34(5):75-76.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:316.
[7]He J,Liao T,Zhong G X,et al.Alzheimer's disease-like early-phase brain pathogenesis:self-curing amelioration of neurodegeneration from pro-inflammatory'wounding'to anti-inflammatory'healing'[J].Curr Alzheimer Res,2017,14(10):1123-1135.
[8]王雪岩,周震,张琳琳,等.淫羊藿苷对SAMP8小鼠记忆能力、神经元微观形态及GSK-3β蛋白表达的影响[J].辽宁中医杂志,2014,41(12):2707-2710.
[9]Wu G,Li L,Li H,et al.Electroacupuncture ameliorates spatial learning and memory impairment via attenuating NOX2-related oxidative stress in a rat model of Alzheimer's disease induced by Aβ1-42[J].Mol Cell Biol,2017,63(4):38.
[10]Cao Y,Zhang L W,Wang J,et al.Mechanisms of acupuncture effect on Alzheimer's disease in animal-based researches[J].Curr Top Med Chem,2016,16(5):574-578.
[11]Mehta D C,Short J L,Nicolazzo J A.Reduced CNS exposure of me-mantine in a triple transgenic mouse model of Alzheimer's disease as-sessed using a novel LC-MS technique[J].Pharmaceut Biomed,2013,85:198-206.
[12]Zhang X,Li G,Guo L,et al.Age-related alteration in cerebral blood flow and energy failure is correlated with cognitive impairment in the senescence-accelerated prone mouse strain 8(SAMP8)[J].Neurol Sci,2013,34(11):1917-1924.
[13]Xiong H,Callaghan D,Jones A,et al.ABCG2 is upregulated in Al-zheimer's brain with cerebral amyloid angiopathy and may act as a gate-keeper at the blood-brain barrier for Aβ1-40 peptides[J].J Neurosci,2009,29(17):5463-5475.
[14]Di Marco L Y,Venneri A,Farkas E,et al.Vascular dysfunction in the pathogenesis of Alzheimer's disease-a review of endothelium-mediat-ed mechanisms and ensuing vicious circles[J].Neurobiol Dis,2015,82:593-606.
[15]Zlokovic B V.The blood-brain barrier in health and chronic neurode-generative disorders[J].Neuron,2008,57(2):178-201.
[16]Butterfield D A,Poon H F.The senescence-accelerated prone mouse(SAMP8):a model of age-related cognitive decline with relevance to al-terations of the gene expression and protein abnormalities in Alzheim-er's disease[J].Exp Gerontol,2005,40(10):774-783.
[17]Del Valle J,Duran-Vilaregut J,Manich G,et al.Cerebral amyloid an-giopathy,blood-brain barrier disruption and amyloid accumulation in SAMP8 mice[J].Neurodegener Dis,2011,8(6):421-429.
[18]杨清华,郭玲,陈清,等.电针刺激对APP/PS1双转基因阿尔兹海默病模型小鼠皮质老年斑形成的影响及机制研究[J].中国康复医学杂志,2018,33(7):770-776.
[19]Ning D,Jing J,Menghan L,et al.Manual acupuncture suppresses the expression of proinflammatory proteins associated with the NLRP3inflammasome in the hippocampus of SAMP8 mice[J].Evid Based Compl Alt,2017,2017:1-8.
[20]Reinhold A,Rittner H.Barrier function in the peripheral and central nervous system-a review[J].Eur J Appl Physiol,2016,469(1):1-12.
[21]廖华宁,汪宁,王艳.紧密连接蛋白与血脑屏障[J].安徽医药,2012,16(3):377-379.
[22]吴丽文,尹飞,彭镜,等.紧密连接蛋白ZO-1,occludin和actin参与缺氧缺血诱导的血脑屏障通透性增加[J].中国当代儿科杂志,2008,10(4):513-516.
[23]Gonzalez-Velasquez F J,Kotarek J A,Moss M A.Soluble aggregates of the amyloid-beta protein selectively stimulate permeability in human brain microvascular endothelial monolayers[J].J Neurochem,2008,107(2):466-477.
[24]Tai L M,Holloway K A,Male D K,et al.Amyloid-beta-induced oc-cludin down-regulation and increased permeability in human brain en-dothelial cells is mediated by MAPK activation[J].Cell Mol Med,2010,14(5):1101-1112.
[25]Gustafsson S,Gustavsson T,Roshanbin S,et al.Blood-brain barrier integrity in a mouse model of Alzheimer's disease with or without acute3D6 immunotherapy[J].Neuropharmacology,2018,143:1-9.
[26]Greta H,Stina S,Fang X T,et al.Bivalent brain shuttle increases an-tibody uptake by monovalent binding to the transferrin receptor[J].Theranostics,2017,7(2):308-318.
[27]Jung Y S,Lee S W,Park J H,et al.Electroacupuncture precondition-ing reduces ROS generation with NOX4 down-regulation and amelio-rates blood-brain barrier disruption after ischemic stroke[J].Biomed Sci,2016,23(1):32.
[28]孔海龙,韦旭,周倩,等.针药结合对阿尔茨海默病模型大鼠海马CA1区ChAT、AChE表达的影响[J].皖南医学院学报,2018,37(4):310-313.