跑台运动改善海马线粒体功能提高APP/PS1小鼠学习记忆能力
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摘要
为研究跑台运动对APP/PS1小鼠海马线粒体融合、分裂作用的影响,将遗传背景为C57BL/6的3月龄APP/PS1小鼠和野生小鼠各42只分别随机分为APP/PS1安静对照组(ADC,n=21)和运动组(ADE,n=21),野生安静对照组(WTC,n=21)和运动组(WTE,n=21)。ADE、WTE组进行12周跑台运动,同时ADC、WTC组置于安静跑台环境。水迷宫实验检测小鼠的空间学习记忆能力,RT-PCR法检测线粒体功能关键酶的mRNA水平,Western印迹检测海马融合、分裂及线粒体关键酶的蛋白质表达情况,透射电镜观察海马线粒体融合、分裂状态。结果发现,6月龄APP/PS1小鼠学习记忆能力降低(P<0. 05);海马线粒体融合蛋白质Mfn1、Mfn2、Opa1表达降低(P<0. 05),线粒体分裂蛋白质Drp1、Mff表达增高(P<0. 05);线粒体膜结构模糊,嵴不明显,线粒体碎片增多,空泡化线粒体增多;线粒体呼吸关键酶COX IV及ATP合酶表达均下调(P<0. 05)。12周跑台运动可逆转APP/PS1小鼠的上述变化,改善海马线粒体结构和功能,提高学习记忆能力。综上提示:12周跑台运动改善APP/PS1小鼠学习记忆能力的机制可能与其对线粒体结构与功能的改善有关。
To study the effect of treadmill exercise on mitochondrial fission and fusion in hippocampus of APP/PS1 mice,thirty three-month-old C57 BL/6 APP/PS1 mice and 30 wild mice were randomly divided into quiet control group( ADC,n = 15) and exercise group( ADE,n = 15) of APP/PS1,and quiet control group( WTC,n = 15) and exercise group( WTE,n = 15) of wild. The ADE and WTE groups performed treadmill exercise for 12 weeks,while the ADC and WTC groups were placed in quiet treadmill environment. Water maze test was used to detect spatial learning and memory ability of mice. RT-PCR was used to detect the mRNA of key enzymes in mitochondrial function. Western blot test was used to detect hippocampal protein expression of fission, fusion and the key enzymes in mitochondria.Transmission electron microscopy was used to observe mitochondrial fission and fusion in hippocampus.The results showed that the learning and memory abilities of 6-month-old APP/PS1 mice decreased( P<0. 05). The expression of Mfn1,Mfn2 and Opa1 in hippocampus decreased( P < 0. 05),and the expression of mitochondrial cleavage proteins Drp1 and Mff increased( P < 0. 05). The structure of mitochondrial membrane was blurred,and crest was not obvious. The mitochondrial fragments increased and vacuolated mitochondria increased. The expression levels of COX IV and ATP synthase were downregulated( P < 0. 05). Treadmill exercise for 12-week could reverse the above changes in APP/PS1 mice,improve the structure and function of hippocampal mitochondria,and improve learning and memory ability. In conclusion,the mechanism of 12-week treadmill exercise improving the learning and memory ability of APP/PS1 mice may be related to the improvement of mitochondrial structure and function.
To study the effect of treadmill exercise on mitochondrial fission and fusion in hippocampus of APP/PS1 mice,thirty three-month-old C57 BL/6 APP/PS1 mice and 30 wild mice were randomly divided into quiet control group( ADC,n = 15) and exercise group( ADE,n = 15) of APP/PS1,and quiet control group( WTC,n = 15) and exercise group( WTE,n = 15) of wild. The ADE and WTE groups performed treadmill exercise for 12 weeks,while the ADC and WTC groups were placed in quiet treadmill environment. Water maze test was used to detect spatial learning and memory ability of mice. RT-PCR was used to detect the mRNA of key enzymes in mitochondrial function. Western blot test was used to detect hippocampal protein expression of fission, fusion and the key enzymes in mitochondria.Transmission electron microscopy was used to observe mitochondrial fission and fusion in hippocampus.The results showed that the learning and memory abilities of 6-month-old APP/PS1 mice decreased( P<0. 05). The expression of Mfn1,Mfn2 and Opa1 in hippocampus decreased( P < 0. 05),and the expression of mitochondrial cleavage proteins Drp1 and Mff increased( P < 0. 05). The structure of mitochondrial membrane was blurred,and crest was not obvious. The mitochondrial fragments increased and vacuolated mitochondria increased. The expression levels of COX IV and ATP synthase were downregulated( P < 0. 05). Treadmill exercise for 12-week could reverse the above changes in APP/PS1 mice,improve the structure and function of hippocampal mitochondria,and improve learning and memory ability. In conclusion,the mechanism of 12-week treadmill exercise improving the learning and memory ability of APP/PS1 mice may be related to the improvement of mitochondrial structure and function.
引文
[1]Manczak M,Calkins M J,Reddy P H.Impaired mitochondrial dynamics and abnormal interaction of amyloid beta with mitochondrial protein Drp1 in neurons from patients with Alzheimer’s disease:implications for neuronal damage[J].Hum Mol Genet,2011,20(13):2495-2509
[2]Manczak M,Reddy P H.Abnormal interaction between the mitochondrial fission protein Drp1 and hyperphosphorylated tau in Alzheimer’s disease neurons:implications for mitochondrial dysfunction and neuronal damage[J].Hum Mol Genet,2012,21(11):2538-2547
[3]Qi X,Qvit N,Su Y C,et al.A novel Drp1 inhibitor diminishes aberrant mitochondrial fission and neurotoxicity[J].J Cell Sci,2013,126(Pt 3):789-802
[4]Xie N,Wang C,Lian Y,et al.Inhibition of mitochondrial fission attenuates Abeta-induced microglia apoptosis[J].Neuroscience,2014,256:36-42
[5]Chen H,Mc Caffery J M,Chan D C.Mitochondrial fusion protects against neurodegeneration in the cerebellum[J].Cell,2007,130(3):548-562
[6]de Souto Barreto P,Andrieu S,Rolland Y,et al.Physical activity domains and cognitive function over three years in older adults with subjective memory complaints:Secondary analysis from the MAPT trial[J].J Sci Med Sport,2018,21(1):52-57
[7]Best J R,Eng J J,Davis J C,et al.Study protocol for Vitality:a proof-of-concept randomised controlled trial of exercise training or complex mental and social activities to promote cognition in adults with chronic stroke[J].BMJ Open,2018,8(3):e021490
[8]Masley S C,Roetzheim R,Clayton G,et al.Lifestyle markers predict cognitive function[J].J Am Coll Nutr,2017,36(8):617-623
[9]Zhou C N,Chao F L,Zhang Y,et al.Sex differences in the white matter and myelinated fibers of APP/PS1 mice and the effects of running exercise on the sex differences of AD mice[J].Front Aging Neurosci,2018,10:243
[10]Alkhouli M F,Hung J,Squire M,et al.Exercise and resveratrol increase fracture resistance in the 3xTg-AD mouse model of Alzheimer’s disease[J].BMC Complement Altern Med,2019,19(1):39
[11]Bo H,Kang W,Jiang N,et al.Exercise-induced neuroprotection of hippocampus in APP/PS1 transgenic mice via upregulation of mitochondrial 8-oxoguanine DNA glycosylase[J].Oxid Med Cell Longev,2014,2014:834502
[12]Baker E J,Gleeson T T.The effects of intensity on the energetics of brief locomotor activity[J].J Exp Biol,1999,202(Pt 22):3081-3087
[13]闫清伟,色里玛,巴桑次仁,等.藏药七十味珍珠丸改善APP/PS1小鼠学习记忆能力[J].中国生物化学与分子生物学报(Yan QW,Se LM,BASang CR,et al.Tibetan medicine RNSPimproves APP/PS1 mice learning and memory ability[J].Chin JBiochem Mol Biol),2017,33(12):1251-1257
[14]Silva D F,Selfridge J E,Lu J,et al.Mitochondrial abnormalities in Alzheimer’s disease:possible targets for therapeutic intervention[J].Adv Pharmacol,2012,64:83-126
[15]Manczak M,Reddy P H.Abnormal interaction between the mitochondrial fission protein Drp1 and hyperphosphorylated tau in Alzheimer’s disease neurons:implications for mitochondrial dysfunction and neuronal damage[J].Hum Mol Genet,2012,21(11):2538-2547
[16]Manczak M,Calkins M J,Reddy P H.Impaired mitochondrial dynamics and abnormal interaction of amyloid beta with mitochondrial protein Drp1 in neurons from patients with Alzheimer’s disease:implications for neuronal damage[J].Hum Mol Genet,2011,20(13):2495-2509
[17]贾振伟.哺乳动物线粒体质量控制的机制[J].中国细胞生物学学报(Jia ZW.The mechanism of mitochondria quality control in mammals[J].Chin J Cell Biol),2017,39(3):373-380
[18]Palmer C S,Osellame L D,Laine D,et al.Mi D49 and Mi D51,new components of the mitochondrial fission machinery[J].EMBO Rep,2011,12(6):565-573
[19]张玉森,董瑞瑞,杨倩,等.阿尔兹海默病中线粒体损伤机制及多酚对其防治的研究进展[J].现代预防医学,(Zhang YuSen,Dong Rui-Rui,Yang Qian,et al.Progress of study on mitochondrial damage mechanisms in Alzheimer’s disease and polyphenol prevention and treatment effects[J].Mod Prev Med),2017,44(13):2411-2414
[20]杨瀚宇,王璐,臧彩霞,等.线粒体动力学调控的分子机制及其与β淀粉样蛋白的关系[J].中国新药杂志(Yang Han-Yu,Wang Lu,Zang Cai-Xia,et al.Molecular mechanism of mitochondrial dynamics regulation and its relationship withβ-amyloid[J].Chin J New Drugs),2017,2017(19):2285-2290
[21]Roy M,Reddy P H,Iijima M,et al.Mitochondrial division and fusion in metabolism[J].Curr Opin Cell Biol,2015,33:111-118
[22]Santarelli R,Rossi R,Scimemi P,et al.OPA1-related auditory neuropathy:site of lesion and outcome of cochlear implantation[J].Brain,2015,138(Pt 3):563-576
[23]Patten D A,Wong J,Khacho M,et al.OPA1-dependent cristae modulation is essential for cellular adaptation to metabolic demand[J].EMBO J,2014,33(22):2676-2691
[24]Anand R,Wai T,Baker M J,et al.The i-AAA protease YME1Land OMA1 cleave OPA1 to balance mitochondrial fusion and fission[J].J Cell Biol,2014,204(6):919-929
[25]Kang S,Moon N R,Kim D S,et al.Central acylated ghrelin improves memory function and hippocampal AMPK activation and partly reverses the impairment of energy and glucose metabolism in rats infused with beta-amyloid[J].Peptides,2015,71:84-93
[26]Pei L,Mu Y,Leblanc M,et al.Dependence of hippocampal function on ERR gamma-regulated mitochondrial metabolism[J].Cell Metab,2015,21(4):628-636
[27]Xia C,Zhu L,Shao W,et al.The Effect of Hippocampal Cognitive Impairment and XIAP on Glucose and Lipids Metabolism in Rats[J].Cell Physiol Biochem,2016,38(2):609-618
[28]Huang T T,Leu D,Zou Y.Oxidative stress and redox regulation on hippocampal-dependent cognitive functions[J].Arch Biochem Biophys,2015,576:2-7
[29]Hernandez-Zimbron L F,Rivas-Arancibia S.Oxidative stress caused by ozone exposure induces beta-amyloid 1-42overproduction and mitochondrial accumulation by activating the amyloidogenic pathway[J].Neuroscience,2015,304:340-348
[30]曹玲芳,李怡芳,何蓉蓉,等.细胞色素c氧化酶对衰老调控作用的研究进展[J].国际药学研究杂志(Cao Ling-Fang,Li Yi-Fang,He Rong-Rong,et al.Regulation function of cytochrome c oxidase on aging:research advances[J].J Int Pharm Res),2013,40(6):751-756,777
[31]Bernstein C,Facista A,Nguyen H,et al.Cancer and age related colonic crypt deficiencies in cytochrome c oxidase I[J].World JGastrointest Oncol,2010,2(12):429-442
[32]Perez-Gracia E,Torrejon-Escribano B,Ferrer I.Dystrophic neurites of senile plaques in Alzheimer’s disease are deficient in cytochrome c oxidase[J].Acta Neuropathol,2008,116(3):261-268
[33]Bosetti F,Brizzi F,Barogi S,et al.Cytochrome c oxidase and mitochondrial F1F0-ATPase(ATP synthase)activities in platelets and brain from patients with Alzheimer’s disease[J].Neurobiol Aging,2002,23(3):371-376
[34]Costa R O,Ferreiro E,Oliveira C R,et al.Inhibition of mitochondrial cytochrome c oxidase potentiates Abeta-induced ERstress and cell death in cortical neurons[J].Mol Cell Neurosci,2013,52:1-8
[35]高静,陈雯,张同霞,等.颞叶癫痫大鼠海马线粒体细胞色素氧化酶亚基Ⅲ和Ⅳ表达的变化[J].山东大学学报(医学版)(Gao Jing,Chen Wen,Zhang Tong-Xia,et al.Changes of expression of mitochondrial cytochrome oxidase subunitsⅢandⅣin the rat hippocampus of temporal lobe epilepsy[J].J Shandong Univ(Health Sci),2007,45(8):817-820,824
[36]Li Y,Park J S,Deng J H,et al.Cytochrome c oxidase subunit IV is essential for assembly and respiratory function of the enzyme complex[J].J Bioenerg Biomembr,2006,38(5-6):283-291
[37]刘道波,刘兴,王志强,等.三磷酸腺苷(ATP)合成酶抑制剂的研究进展[J].沈阳药科大学学报(Liu DB,Liu X,Wang ZQ,et al.Progress in study of ATP synthase inhibitors[J].JShenyang Pharm Univ),2012,2012(11):872-886
[38]Zhao F L,Fang F,Qiao P F,et al.AP39,a Mitochondriatargeted hydrogen sulfide donor,supports cellular bioenergetics and protects against Alzheimer’s disease by preserving mitochondrial function in APP/PS1 mice and neurons[J].Oxid Med Cell Longev,2016,2016:8360738
[2]Manczak M,Reddy P H.Abnormal interaction between the mitochondrial fission protein Drp1 and hyperphosphorylated tau in Alzheimer’s disease neurons:implications for mitochondrial dysfunction and neuronal damage[J].Hum Mol Genet,2012,21(11):2538-2547
[3]Qi X,Qvit N,Su Y C,et al.A novel Drp1 inhibitor diminishes aberrant mitochondrial fission and neurotoxicity[J].J Cell Sci,2013,126(Pt 3):789-802
[4]Xie N,Wang C,Lian Y,et al.Inhibition of mitochondrial fission attenuates Abeta-induced microglia apoptosis[J].Neuroscience,2014,256:36-42
[5]Chen H,Mc Caffery J M,Chan D C.Mitochondrial fusion protects against neurodegeneration in the cerebellum[J].Cell,2007,130(3):548-562
[6]de Souto Barreto P,Andrieu S,Rolland Y,et al.Physical activity domains and cognitive function over three years in older adults with subjective memory complaints:Secondary analysis from the MAPT trial[J].J Sci Med Sport,2018,21(1):52-57
[7]Best J R,Eng J J,Davis J C,et al.Study protocol for Vitality:a proof-of-concept randomised controlled trial of exercise training or complex mental and social activities to promote cognition in adults with chronic stroke[J].BMJ Open,2018,8(3):e021490
[8]Masley S C,Roetzheim R,Clayton G,et al.Lifestyle markers predict cognitive function[J].J Am Coll Nutr,2017,36(8):617-623
[9]Zhou C N,Chao F L,Zhang Y,et al.Sex differences in the white matter and myelinated fibers of APP/PS1 mice and the effects of running exercise on the sex differences of AD mice[J].Front Aging Neurosci,2018,10:243
[10]Alkhouli M F,Hung J,Squire M,et al.Exercise and resveratrol increase fracture resistance in the 3xTg-AD mouse model of Alzheimer’s disease[J].BMC Complement Altern Med,2019,19(1):39
[11]Bo H,Kang W,Jiang N,et al.Exercise-induced neuroprotection of hippocampus in APP/PS1 transgenic mice via upregulation of mitochondrial 8-oxoguanine DNA glycosylase[J].Oxid Med Cell Longev,2014,2014:834502
[12]Baker E J,Gleeson T T.The effects of intensity on the energetics of brief locomotor activity[J].J Exp Biol,1999,202(Pt 22):3081-3087
[13]闫清伟,色里玛,巴桑次仁,等.藏药七十味珍珠丸改善APP/PS1小鼠学习记忆能力[J].中国生物化学与分子生物学报(Yan QW,Se LM,BASang CR,et al.Tibetan medicine RNSPimproves APP/PS1 mice learning and memory ability[J].Chin JBiochem Mol Biol),2017,33(12):1251-1257
[14]Silva D F,Selfridge J E,Lu J,et al.Mitochondrial abnormalities in Alzheimer’s disease:possible targets for therapeutic intervention[J].Adv Pharmacol,2012,64:83-126
[15]Manczak M,Reddy P H.Abnormal interaction between the mitochondrial fission protein Drp1 and hyperphosphorylated tau in Alzheimer’s disease neurons:implications for mitochondrial dysfunction and neuronal damage[J].Hum Mol Genet,2012,21(11):2538-2547
[16]Manczak M,Calkins M J,Reddy P H.Impaired mitochondrial dynamics and abnormal interaction of amyloid beta with mitochondrial protein Drp1 in neurons from patients with Alzheimer’s disease:implications for neuronal damage[J].Hum Mol Genet,2011,20(13):2495-2509
[17]贾振伟.哺乳动物线粒体质量控制的机制[J].中国细胞生物学学报(Jia ZW.The mechanism of mitochondria quality control in mammals[J].Chin J Cell Biol),2017,39(3):373-380
[18]Palmer C S,Osellame L D,Laine D,et al.Mi D49 and Mi D51,new components of the mitochondrial fission machinery[J].EMBO Rep,2011,12(6):565-573
[19]张玉森,董瑞瑞,杨倩,等.阿尔兹海默病中线粒体损伤机制及多酚对其防治的研究进展[J].现代预防医学,(Zhang YuSen,Dong Rui-Rui,Yang Qian,et al.Progress of study on mitochondrial damage mechanisms in Alzheimer’s disease and polyphenol prevention and treatment effects[J].Mod Prev Med),2017,44(13):2411-2414
[20]杨瀚宇,王璐,臧彩霞,等.线粒体动力学调控的分子机制及其与β淀粉样蛋白的关系[J].中国新药杂志(Yang Han-Yu,Wang Lu,Zang Cai-Xia,et al.Molecular mechanism of mitochondrial dynamics regulation and its relationship withβ-amyloid[J].Chin J New Drugs),2017,2017(19):2285-2290
[21]Roy M,Reddy P H,Iijima M,et al.Mitochondrial division and fusion in metabolism[J].Curr Opin Cell Biol,2015,33:111-118
[22]Santarelli R,Rossi R,Scimemi P,et al.OPA1-related auditory neuropathy:site of lesion and outcome of cochlear implantation[J].Brain,2015,138(Pt 3):563-576
[23]Patten D A,Wong J,Khacho M,et al.OPA1-dependent cristae modulation is essential for cellular adaptation to metabolic demand[J].EMBO J,2014,33(22):2676-2691
[24]Anand R,Wai T,Baker M J,et al.The i-AAA protease YME1Land OMA1 cleave OPA1 to balance mitochondrial fusion and fission[J].J Cell Biol,2014,204(6):919-929
[25]Kang S,Moon N R,Kim D S,et al.Central acylated ghrelin improves memory function and hippocampal AMPK activation and partly reverses the impairment of energy and glucose metabolism in rats infused with beta-amyloid[J].Peptides,2015,71:84-93
[26]Pei L,Mu Y,Leblanc M,et al.Dependence of hippocampal function on ERR gamma-regulated mitochondrial metabolism[J].Cell Metab,2015,21(4):628-636
[27]Xia C,Zhu L,Shao W,et al.The Effect of Hippocampal Cognitive Impairment and XIAP on Glucose and Lipids Metabolism in Rats[J].Cell Physiol Biochem,2016,38(2):609-618
[28]Huang T T,Leu D,Zou Y.Oxidative stress and redox regulation on hippocampal-dependent cognitive functions[J].Arch Biochem Biophys,2015,576:2-7
[29]Hernandez-Zimbron L F,Rivas-Arancibia S.Oxidative stress caused by ozone exposure induces beta-amyloid 1-42overproduction and mitochondrial accumulation by activating the amyloidogenic pathway[J].Neuroscience,2015,304:340-348
[30]曹玲芳,李怡芳,何蓉蓉,等.细胞色素c氧化酶对衰老调控作用的研究进展[J].国际药学研究杂志(Cao Ling-Fang,Li Yi-Fang,He Rong-Rong,et al.Regulation function of cytochrome c oxidase on aging:research advances[J].J Int Pharm Res),2013,40(6):751-756,777
[31]Bernstein C,Facista A,Nguyen H,et al.Cancer and age related colonic crypt deficiencies in cytochrome c oxidase I[J].World JGastrointest Oncol,2010,2(12):429-442
[32]Perez-Gracia E,Torrejon-Escribano B,Ferrer I.Dystrophic neurites of senile plaques in Alzheimer’s disease are deficient in cytochrome c oxidase[J].Acta Neuropathol,2008,116(3):261-268
[33]Bosetti F,Brizzi F,Barogi S,et al.Cytochrome c oxidase and mitochondrial F1F0-ATPase(ATP synthase)activities in platelets and brain from patients with Alzheimer’s disease[J].Neurobiol Aging,2002,23(3):371-376
[34]Costa R O,Ferreiro E,Oliveira C R,et al.Inhibition of mitochondrial cytochrome c oxidase potentiates Abeta-induced ERstress and cell death in cortical neurons[J].Mol Cell Neurosci,2013,52:1-8
[35]高静,陈雯,张同霞,等.颞叶癫痫大鼠海马线粒体细胞色素氧化酶亚基Ⅲ和Ⅳ表达的变化[J].山东大学学报(医学版)(Gao Jing,Chen Wen,Zhang Tong-Xia,et al.Changes of expression of mitochondrial cytochrome oxidase subunitsⅢandⅣin the rat hippocampus of temporal lobe epilepsy[J].J Shandong Univ(Health Sci),2007,45(8):817-820,824
[36]Li Y,Park J S,Deng J H,et al.Cytochrome c oxidase subunit IV is essential for assembly and respiratory function of the enzyme complex[J].J Bioenerg Biomembr,2006,38(5-6):283-291
[37]刘道波,刘兴,王志强,等.三磷酸腺苷(ATP)合成酶抑制剂的研究进展[J].沈阳药科大学学报(Liu DB,Liu X,Wang ZQ,et al.Progress in study of ATP synthase inhibitors[J].JShenyang Pharm Univ),2012,2012(11):872-886
[38]Zhao F L,Fang F,Qiao P F,et al.AP39,a Mitochondriatargeted hydrogen sulfide donor,supports cellular bioenergetics and protects against Alzheimer’s disease by preserving mitochondrial function in APP/PS1 mice and neurons[J].Oxid Med Cell Longev,2016,2016:8360738