有氧运动改善AD模型APP/PS1双转基因小鼠中枢神经元线粒体融合分裂动态平衡
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摘要
线粒体融合分裂平衡是线粒体动力学的需要。本研究观察12周规律有氧运动对APP/PS1双转基因小鼠中枢神经元线粒体融合分裂动态平衡的影响。本研究采用3月龄雄性APP/PS1小鼠(AD模型)随机分为AD安静组(AS)、AD运动组(AE),同月龄雄性C57BL/6J小鼠做正常对照组(CS)。AE组进行12周规律跑台运动,5 d/周,60 min/d。前10 min运动速度12 m/min,后50min运动速度15 m/min,跑台坡度为0°。八臂迷宫实验检测小鼠工作记忆错误频率和参考记忆错误频率; Western印迹检测小鼠皮层、海马组织中线粒体分裂蛋白Drp1和Fis1的含量,以及Drp1的活性(p-Drp1-Ser616)、线粒体融合蛋白Mfn1、Mfn2、Opa1的表达水平;透射电镜观察皮层、海马线粒体形态结构、健康线粒体比率及线粒体平均直径。本研究证实AS组较CS组工作记忆错误频率显著提高(P<0. 05),12周有氧运动显著降低工作记忆错误频率(P<0. 05)。AS组小鼠皮层Fis1蛋白和海马脑区Drp1、Fis1蛋白表达水平及皮层、海马脑区Drp1蛋白的活性增加(P<0. 05)。而皮层Mfn1和海马Mfn1、Mfn2蛋白表达水平显著降低(P<0. 05)。12周有氧运动显著减低Fis1、Drp1蛋白表达及Drp1蛋白的活性,提高Mfn1、Mfn2蛋白表达水平(P<0. 05)。AS组小鼠皮层、海马线粒体多呈现球形,部分线粒体膜结构消失,线粒体嵴结构紊乱。且AS组较CS组小鼠健康线粒体比率降低、直径缩短。12周规律有氧运动可明显改善线粒体形态和结构,提高健康线粒体比率及直径。本研究提示,12周规律有氧运动可有效抑制皮层、海马脑区线粒体分裂蛋白Drp1和Fis1的表达,降低Drp1的活性(p-Drp1-Ser616),上调线粒体融合蛋白Mfn1、Mfn2的蛋白表达水平,改善线粒体形态和结构以促进线粒体质量控制,是有氧运动改善AD模型空间学习记忆能力的分子机制之一。
Mitochondrial dynamics,including mitochondrial fusion and fission balance,is essential for the mitochondria quality control. This study observed the effects of 12 weeks aerobic exercise onmitochondrial dynamics in 6-month-old APP/PS1 transgenic mice. C57 BL/6 J mice( 3-month-old) and APP/PS1 transgenic mice( 3-month-old) were randomly divided into sedentary( CS,AS) and 12 weeks exercise( AE),respectively. The mice in AE group underwent treadmill at 12 m/min( the first 10 min)or 15 m/min( the following 50 min) for 60 min,0° slope,5 d/week,for 12 weeks. The behavioral changes were detected by eight arm experiments to observe the frequency of errors of working memory and reference memory. The protein levels of dynamin-related protein( Drp1),p-Drp1-Ser616,fission 1 protein( Fis1),mitofusin 1( Mfn1),mitofusin 2( Mfn2),and optic atrophy 1( Opa1) were measured using Western blotting. The mitochondrial morphology, ratio of healthy mitochondrial and average mitochondrial diameter in the brain tissues were detected by transmission electron microscope.Behaviorally: there was significant changes in 6-month-old AS compared with the control cohorts( CS) in the frequency of working memory errors( P < 0. 05). The AE mice performed significantly better than those of AS in the frequency of working memory errors( P<0. 05). The mice of 6 month-old APP/PS1 group displayed a prominent increase in the protein levels of Fis1 in the cortex,and in the hippocampus compared with the CS controls( P < 0. 05). Increase of protein levels and activity of Drp1( p-Drp1-Ser616) was detected in hippocampus. A decrease of protein levels of Mfn1 in the cortex,and Mfn1 and Mfn2 in the hippocampus was also found. Aerobic exercise decreased the expression of Drp1 and Fis1 reduced the activity of Drp1 and increased Mfn1 and Mfn2 protein expression in APP/PS1 mice( P <0. 05). The results of transmission electron showed that the ratio of healthy mitochondrial and average mitochondrial diameter decreased in 6-month-old APP/PS1 mice. Swollen mitochondria and ruptured membranes were also observed. The 12 weeks aerobic exercise increased the ratio of healthy mitochondria and average mitochondrial diameter and improved the mitochondrial structure and function. The results suggest that the 12 weeks aerobic exercise can regulate the mitochondrial structure and function of APP/PS1 mice by significantly reducing the expression of Drp1 and Fis1,and the activity of Drp1,and increasing the expression of Mfn1 and Mfn2. Our study may provide information in understating the molecular mechanism of exercise-induced spatial learning and memory ability.
Mitochondrial dynamics,including mitochondrial fusion and fission balance,is essential for the mitochondria quality control. This study observed the effects of 12 weeks aerobic exercise onmitochondrial dynamics in 6-month-old APP/PS1 transgenic mice. C57 BL/6 J mice( 3-month-old) and APP/PS1 transgenic mice( 3-month-old) were randomly divided into sedentary( CS,AS) and 12 weeks exercise( AE),respectively. The mice in AE group underwent treadmill at 12 m/min( the first 10 min)or 15 m/min( the following 50 min) for 60 min,0° slope,5 d/week,for 12 weeks. The behavioral changes were detected by eight arm experiments to observe the frequency of errors of working memory and reference memory. The protein levels of dynamin-related protein( Drp1),p-Drp1-Ser616,fission 1 protein( Fis1),mitofusin 1( Mfn1),mitofusin 2( Mfn2),and optic atrophy 1( Opa1) were measured using Western blotting. The mitochondrial morphology, ratio of healthy mitochondrial and average mitochondrial diameter in the brain tissues were detected by transmission electron microscope.Behaviorally: there was significant changes in 6-month-old AS compared with the control cohorts( CS) in the frequency of working memory errors( P < 0. 05). The AE mice performed significantly better than those of AS in the frequency of working memory errors( P<0. 05). The mice of 6 month-old APP/PS1 group displayed a prominent increase in the protein levels of Fis1 in the cortex,and in the hippocampus compared with the CS controls( P < 0. 05). Increase of protein levels and activity of Drp1( p-Drp1-Ser616) was detected in hippocampus. A decrease of protein levels of Mfn1 in the cortex,and Mfn1 and Mfn2 in the hippocampus was also found. Aerobic exercise decreased the expression of Drp1 and Fis1 reduced the activity of Drp1 and increased Mfn1 and Mfn2 protein expression in APP/PS1 mice( P <0. 05). The results of transmission electron showed that the ratio of healthy mitochondrial and average mitochondrial diameter decreased in 6-month-old APP/PS1 mice. Swollen mitochondria and ruptured membranes were also observed. The 12 weeks aerobic exercise increased the ratio of healthy mitochondria and average mitochondrial diameter and improved the mitochondrial structure and function. The results suggest that the 12 weeks aerobic exercise can regulate the mitochondrial structure and function of APP/PS1 mice by significantly reducing the expression of Drp1 and Fis1,and the activity of Drp1,and increasing the expression of Mfn1 and Mfn2. Our study may provide information in understating the molecular mechanism of exercise-induced spatial learning and memory ability.
引文
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[10]于晓伟,解玉珍,顾博雅,等.有氧运动调节驱动蛋白改善AD模型皮层线粒体轴突转运[J].北京体育大学学报(Yu XW,Xie YZ,Gu BY,et al. Effects of aerobic exercise on Kinesin improving the axonal mitochondrial transport[J]. J Beijing Sport Univ),2016,39(6):63-68
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[14] Stockburger C, Miano D, Pallas T, et al. Enhanced neuroplasticity by the metabolic enhancer piracetam associated with improved mitochondrial dynamics and altered permeability transition pore function[J]. Neural Plast,2016,2016:8075903
[15] Fu L, Sun Y, Guo Y, et al. Progresscive spatial memory impairment, brain amyloid deposition and changes in serum amyloid levels as a function of age in APPswe/PS1dE9 mice[J].Curr Alzheimer Res,2018,15(11):1053-1061
[16]於来康,顾博雅,李岩,等.有氧运动调节APP/PS1小鼠海马CaMKIIα、AMPAR活性,增加突触可塑性[J].北京体育大学学报(Yu LK,Gu BY,Li Y,et al. Aerobic exercise increase synaptic by regulating the activity of AMPA receptor and CaMK IIαin the hippocampus of APP/PS1 mice[J]. J Beijing Sport Univ),2017,40(1):41-45
[17] Lipaus IFS,Gomes EF,Martins CW,et al. Impairment of spatial working memory and oxidative stress induced by repeated crack cocaine inhalation in rats[J]. Behav Brain Res,2019,359:910-917
[18] Cai Q,Tammineni P. Alterations in mitochondrial quality control in Alzheimer’s disease[J]. Front Cell Neurosci,2016,10:24
[19] Serasinghe MN,Wieder SY,Renault TT,et al. Mitochondrial division is requisite to RAS-induced transformation and targeted by oncogenic MAPK pathway inhibitors[J]. Mol Cell,2015,57(3):521-536
[20] Chandhok G,Lazarou M,Neumann B. Structure,function,and regulation of mitofusin-2 in health and disease[J]. Biol Rev Camb Philos Soc,2018,93(2):933-949
[21] Dhingra R, Kirshenbaum LA. Regulation of mitochondrial dynamics and cell fate[J]. Circ J,2014,78(4):803-810
[22] Yan L,Qi Y,Huang X,et al. Stuctural basis for GTP hydrolysis and conformational change of MFN 1 in mediating membrane fusion[J]. Nat Stuct Mol Biol,2018,25(3):233-243
[23] Cogliati S,Frezza C,Soriano ME,et al. Mitochondrial cristae shape determines respiratory chain supercomplexes assembly and respiratory efficiency[J]. Cell,2013,155(1):160-171
[24] Lee H,Smith SB,Yoon Y. The short variant of the mitochondrial dynamin OPA1 maintains mitochondrial energetic and cristae structure[J]. J Biol Chem,2017,292(17):7115-7130
[25] Mancza M,Kandimalla R,Fry D,et al. Protective effects of reduced dynamin-related protein 1 against amyloid beta induced mitochondrial dysfunction and synaptic damage in Alzheimer’s disease[J]. Hum Mol Genet,2016,25(23):5148-5166
[2] Labbe K,Murley A,Nunnari J. Determinants and functions of mitochondrial behavior[J]. Annu Rev Cell Dev Biol,2014,30:357-391
[3] Otera H,Ishihara N,Mihara K. New insights into the function and regulation of mitochondrial fission[J]. Biochim Biophys Acta,2013,1833(35):1256-1268
[4] Ishihara N,Otera H,Oka T,et al. Regulation and physiologic functions of GTPases in mitochondrial fusion and fission in mammals[J]. Antioxid Redox Signal,2013,19(4):389-399
[5] Romanello V,Sandri M. Mitochondrial quality control and muscle mass maintenance[J]. Front Physiol,2016,6:422
[6] Gao J,Wang L,Liu J,et al. Abnormalities of mitochondrial dynamics in neurodegenerative diseases[J]. Antioxidants(Basel),2017,6(2). pii:E25
[7] Kim DI, Lee KH, Gabr AA, et al. Aβ-induced Drp1phosphorylation through Akt activation promotes excessive tochondrial fission leading to neuronal apoptosis[J]. Biochim Biophys Acta,2016,1863(11):2820-2834
[8] Wang L, Guo L,Lu L, et al. Synaptosomal mitochondrial dysfunction in 5xFAD mouse model of Alzheimer’s disease[J].PLoS One,2016,11(3):e0150441
[9] Correia SC,Perry G,Moreira PI. Mitochondrial traffic jams in Alzheimer’s disease–pinpointing the roadblocks[J]. Biochim Biophys Acta,2016,1862(10):1909-1917
[10]于晓伟,解玉珍,顾博雅,等.有氧运动调节驱动蛋白改善AD模型皮层线粒体轴突转运[J].北京体育大学学报(Yu XW,Xie YZ,Gu BY,et al. Effects of aerobic exercise on Kinesin improving the axonal mitochondrial transport[J]. J Beijing Sport Univ),2016,39(6):63-68
[11] Friedland-Leuner K, Stockburger C, Denzer I, et al.Mitochondrial dysfunction:cause and consequence of Alzheimer’s disease[J]. Prog Mol Biol Transl Sci,2014,127:183-210
[12] Wirz KT,Keitel S,Swaab DF,et al. Early molecular changes in Alzheimer disease:can we catch the disease in its presymptomatic phase?[J]. J Alzheimers Dis,2014,38(4):719-740
[13] Swerdlow RH,Burns JM,Khan SM. The Alzheimer’s disease mitochondrial cascade hypothesis:progress and perspectives[J].Biochim Biophys Acta,2014,1842(8):1219-1231
[14] Stockburger C, Miano D, Pallas T, et al. Enhanced neuroplasticity by the metabolic enhancer piracetam associated with improved mitochondrial dynamics and altered permeability transition pore function[J]. Neural Plast,2016,2016:8075903
[15] Fu L, Sun Y, Guo Y, et al. Progresscive spatial memory impairment, brain amyloid deposition and changes in serum amyloid levels as a function of age in APPswe/PS1dE9 mice[J].Curr Alzheimer Res,2018,15(11):1053-1061
[16]於来康,顾博雅,李岩,等.有氧运动调节APP/PS1小鼠海马CaMKIIα、AMPAR活性,增加突触可塑性[J].北京体育大学学报(Yu LK,Gu BY,Li Y,et al. Aerobic exercise increase synaptic by regulating the activity of AMPA receptor and CaMK IIαin the hippocampus of APP/PS1 mice[J]. J Beijing Sport Univ),2017,40(1):41-45
[17] Lipaus IFS,Gomes EF,Martins CW,et al. Impairment of spatial working memory and oxidative stress induced by repeated crack cocaine inhalation in rats[J]. Behav Brain Res,2019,359:910-917
[18] Cai Q,Tammineni P. Alterations in mitochondrial quality control in Alzheimer’s disease[J]. Front Cell Neurosci,2016,10:24
[19] Serasinghe MN,Wieder SY,Renault TT,et al. Mitochondrial division is requisite to RAS-induced transformation and targeted by oncogenic MAPK pathway inhibitors[J]. Mol Cell,2015,57(3):521-536
[20] Chandhok G,Lazarou M,Neumann B. Structure,function,and regulation of mitofusin-2 in health and disease[J]. Biol Rev Camb Philos Soc,2018,93(2):933-949
[21] Dhingra R, Kirshenbaum LA. Regulation of mitochondrial dynamics and cell fate[J]. Circ J,2014,78(4):803-810
[22] Yan L,Qi Y,Huang X,et al. Stuctural basis for GTP hydrolysis and conformational change of MFN 1 in mediating membrane fusion[J]. Nat Stuct Mol Biol,2018,25(3):233-243
[23] Cogliati S,Frezza C,Soriano ME,et al. Mitochondrial cristae shape determines respiratory chain supercomplexes assembly and respiratory efficiency[J]. Cell,2013,155(1):160-171
[24] Lee H,Smith SB,Yoon Y. The short variant of the mitochondrial dynamin OPA1 maintains mitochondrial energetic and cristae structure[J]. J Biol Chem,2017,292(17):7115-7130
[25] Mancza M,Kandimalla R,Fry D,et al. Protective effects of reduced dynamin-related protein 1 against amyloid beta induced mitochondrial dysfunction and synaptic damage in Alzheimer’s disease[J]. Hum Mol Genet,2016,25(23):5148-5166