运动上调蛋白酶体活性促进SVZ区神经发生
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摘要
目的:观察运动后小鼠脑室室管膜下区(subventricular zone,SVZ)蛋白酶体活性变化,探讨运动促进神经发生的机制。方法:应用荧光分光光度法检测新生小鼠(Postnantal day 0,P0)、成年小鼠(Postnantal day 90,P90)及老年小鼠(Postnantal day 540,P540)SVZ蛋白酶体活性。成年小鼠自主跑轮运动4周后检测蛋白酶体活性,通过Ki67、DCX免疫荧光染色检测SVZ神经发生水平;小鼠注射MG132,观察抑制蛋白酶体活性是否影响运动对神经发生的促进作用。结果:随年龄增长,P90、P540小鼠SVZ蛋白酶体活性较P0小鼠分别下降50.9%(P<0.01)、70.4%(P<0.01)。运动后小鼠SVZ蛋白酶体活性较安静组增加39.8%(P<0.01),蛋白酶体亚单位PSMB1、PSMB2、PSMB5及PSMA3的表达水平也较安静组分别增加9.1%(P<0.05)、21.7%(P<0.01)、25.4%(P<0.01)和10.1%(P<0.05),神经发生相关指标Ki67~+、DCX~+细胞数量较安静组分别增加32.4%(P<0.01)和78.3%(P<0.001)。小鼠侧脑室注射MG132后再进行跑轮运动,MG132-运动组SVZ区Ki67~+、DCX~+细胞数量较DMSO-运动组分别下降27.6%(P<0.001)和51.5%(P<0.001)。结论:随年龄增长,蛋白酶体活性下降。运动可能通过提高SVZ蛋白酶体活性促进神经发生。
Objective To observe the proteasome activity of physical running mice to clarify the mechanism of its promoting the neurogenesis of the subventricular zone(SVZ). Methods SVZ tissues of newborn mie(Postnatal day 0,P0),adult mice(Postnatal day 90,P90) and aged mice(Postnatal day 540, P540) were isolated to detect the proteasome activity using the fluorescence spectrophotometry.The P90 mice were selected to establish a voluntary running wheel model,and the change of SVZ pro?teasome activity was observed after running exercises for 4 weeks,then Ki67 and DCX immunofluo?rescene staining was used to observe the SVZ neurogenesis of adult mice. The proteasome inhibitor MG132 was injected into SVZ to observe the neurogenesis level of physical running mice. Results The proteasome activity of SVZ decreased gradually with age,and that of P90 and P540 mice decreased50.9%(P<0.01) and 70.4%(P<0.01) respectively compared with P0 mice. After running for 4 weeks,the proteasome activity of SVZ in P90 mice increased 39.8% compared with those sedentary mice(P<0.01),and the expression of the subunit of proteasome including PSMB1,PSMB2,PSMB5 and PS?MA3 of the running mice increased 9.1%(P<0.05),21.7%(P<0.01),25.4%(P<0.01) and 10.1%(P<0.05) compared to those sedentary ones. The number of the marker of neurogenesis including Ki67 and DCX increased 32.4%(P<0.01) and 78.3%(P<0.001) respectively compared to the sedentary mice.After injecting MG132,the number of Ki67 and DCX of the running mice decreased 27.6%(P<0.001)and 51.5%(P<0.001). Conclusion The proteasome activity of SVZ decreased with age. Physical running may promote neurogenesis by increasing the proteasome avtivity of SVZ.
Objective To observe the proteasome activity of physical running mice to clarify the mechanism of its promoting the neurogenesis of the subventricular zone(SVZ). Methods SVZ tissues of newborn mie(Postnatal day 0,P0),adult mice(Postnatal day 90,P90) and aged mice(Postnatal day 540, P540) were isolated to detect the proteasome activity using the fluorescence spectrophotometry.The P90 mice were selected to establish a voluntary running wheel model,and the change of SVZ pro?teasome activity was observed after running exercises for 4 weeks,then Ki67 and DCX immunofluo?rescene staining was used to observe the SVZ neurogenesis of adult mice. The proteasome inhibitor MG132 was injected into SVZ to observe the neurogenesis level of physical running mice. Results The proteasome activity of SVZ decreased gradually with age,and that of P90 and P540 mice decreased50.9%(P<0.01) and 70.4%(P<0.01) respectively compared with P0 mice. After running for 4 weeks,the proteasome activity of SVZ in P90 mice increased 39.8% compared with those sedentary mice(P<0.01),and the expression of the subunit of proteasome including PSMB1,PSMB2,PSMB5 and PS?MA3 of the running mice increased 9.1%(P<0.05),21.7%(P<0.01),25.4%(P<0.01) and 10.1%(P<0.05) compared to those sedentary ones. The number of the marker of neurogenesis including Ki67 and DCX increased 32.4%(P<0.01) and 78.3%(P<0.001) respectively compared to the sedentary mice.After injecting MG132,the number of Ki67 and DCX of the running mice decreased 27.6%(P<0.001)and 51.5%(P<0.001). Conclusion The proteasome activity of SVZ decreased with age. Physical running may promote neurogenesis by increasing the proteasome avtivity of SVZ.
引文
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[14]Saraulli D,Costanzi M,Mastrorilli V,et al.The long run:neuroprotective effects of physical exercise on adult neurogenesis from youth to old age.Curr Neuropharmacol,2017,15(4):519-533.
[15]Mastrorilli V,Scopa C,Saraulli D,et al.Physical exercise rescues defective neural stem cells and neurogenesis in the adult subventricular zone of Btg1 knockout mice.Brain Struct Funct,2017,222(6):2855-2876.
[16]Farioli-Vecchioli S,Mattera A,Micheli L,et al.Running rescues defective adult neurogenesis by shortening the length of the cell cycle of neural stem and progenitor cells.Stem cells,2014,32(7):1968-1982.
[17]Zhang Y,Yang B,Zhao J,et al.Proteasome Inhibitor Carbobenzoxy-L-Leucyl-L-Leucyl-L-Leucinal(MG132)Enhances Therapeutic Effect of Paclitaxel on Breast Cancer by Inhibiting Nuclear Factor(NF)-kappaB Signaling.Med Sci Monit,2018,24:294-304.
[2]Daynac M,Morizur L,Chicheportiche A,et al.Age-related neurogenesis decline in the subventricular zone is associated with specific cell cycle regulation changes in activated neural stem cells.Sci Rep,2016,6:21505.
[3]Farioli-Vecchioli S,Tirone F.Control of the cell cycle in adult neurogenesis and its relation with physical exercise.Brain Plast,2015,1(1):41-54.
[4]Winocur G,Wojtowicz JM,Huang J,et al.Physical exercise prevents suppression of hippocampal neurogenesis and reduces cognitive impairment in chemotherapy-treated rats.Psychopharmacology,2014,231(11):2311-2320.
[5]Zhao Y,Liu X,He Z,et al.Essential role of proteasomes in maintaining self-renewal in neural progenitor cells.Sci Rep,2016,6:19752.
[6]Ben-Nissan G,Sharon M.Regulating the 20S proteasome ubiquitin-independent degradation pathway.Biomolecules,2014,4(3):862-884.
[7]宋慧芳,杨佳超,牛晓洁,等.叔丁基对苯二酚激活骨髓间充质干细胞蛋白酶体活性延缓复制性衰老.中国病理生理杂志,2015,31(9):1647-1651.
[8]张伟,刘雪芹,苗苗,等.20S蛋白酶体β5亚单位基因沉默对人骨髓间充质干细胞增殖能力的影响.解剖学杂志,2015,38(2):153-156.
[9]Zhang W,Gu GJ,Zhang Q,et al.NSCs promote hippocampal neurogenesis,metabolic changes and synaptogenesis in APP/PS1 transgenic mice.Hippocampus,2017,27(12):1250-1263.
[10]杨佳超,赵云鹤,蒋蓉,等.18α-GA通过上调蛋白酶体活性促进晚期骨髓间充质干细胞增殖.中国病理生理杂志,2015,31(12):2183-2187.
[11]金雪佳.泛素-蛋白酶体系统在神经退行性疾病中的研究.药物生物技术,2015(4):366-368.
[12]Saez I,Vilchez D.The Mechanistic Links Between Proteasome Activity,Aging and Age-related Diseases.Curr Genomics,2014,15(1):38-51.
[13]Lee CS,Lee C,Hu T,et al.Loss of nuclear factor E2-related factor 1 in the brain leads to dysregulation of proteasome gene expression and neurodegeneration.Proc Natl Acad Sci U S A,2011,108(20):8408-8413.
[14]Saraulli D,Costanzi M,Mastrorilli V,et al.The long run:neuroprotective effects of physical exercise on adult neurogenesis from youth to old age.Curr Neuropharmacol,2017,15(4):519-533.
[15]Mastrorilli V,Scopa C,Saraulli D,et al.Physical exercise rescues defective neural stem cells and neurogenesis in the adult subventricular zone of Btg1 knockout mice.Brain Struct Funct,2017,222(6):2855-2876.
[16]Farioli-Vecchioli S,Mattera A,Micheli L,et al.Running rescues defective adult neurogenesis by shortening the length of the cell cycle of neural stem and progenitor cells.Stem cells,2014,32(7):1968-1982.
[17]Zhang Y,Yang B,Zhao J,et al.Proteasome Inhibitor Carbobenzoxy-L-Leucyl-L-Leucyl-L-Leucinal(MG132)Enhances Therapeutic Effect of Paclitaxel on Breast Cancer by Inhibiting Nuclear Factor(NF)-kappaB Signaling.Med Sci Monit,2018,24:294-304.