间歇有氧运动通过激活前额叶皮质线粒体自噬改善衰老模型大鼠学习记忆能力  

Aerobic interval exercise improves learning and memory ability of aging model rats by activation of mitophagy in prefrontal cortex

作　　者：张金梅[1,2] 伍怡 刘仁凡 吕丽婷 高巧婧 高骏 李雪 金毓[1] 王璐 ZHANG Jinmei;WU Yi;LIU Renfan;LÜLiting;GAO Qiaojing;GAO Jun;LI Xue;JIN Yu;WANG Lu(Chengdu Sport University,Chengdu 610041,China;Chaohu University,Hefei 238000,China)

机构地区：[1]成都体育学院,四川成都610041 [2]巢湖学院,安徽合肥238000

出　　处：《中国病理生理杂志》2023年第3期400-408,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31900848);四川省科技厅项目基金(No.2022NSFSC1724);运动医学四川省重点实验室暨或国家体育总局运动医学重点实验室资助项目(No.2022-A033);成都体育学院运动医学与健康研究所/郑怀贤骨伤研究所2021-2022年创新课题(No.CX21B05,No.CX21C09,No.CX21A02);安徽省教育厅科学研究重点项目(No.2022AH051691);巢湖学院青少年主动健康智能促进研究中心项目(No.kj22yjzx12)。

摘　　要：目的:探讨间歇有氧运动对D-半乳糖(D-Gal)致衰老模型大鼠前额叶皮质(PFC)线粒体自噬及空间学习记忆能力的影响。方法:8周龄SPF级Sprague-Dawley(SD)大鼠随机分为对照(control)组、衰老(aging)组和aging+运动(Exe)组,每组10只。aging组和aging+Exe组进行6周的D-Gal注射(control组注射等体积的生理盐水);之后aging+Exe组进行8周的间歇有氧运动干预,45 min/d,每周5 d(期间control和aging组自然饲养)。运动干预结束后各组进行Morris水迷宫行为学训练和测试;ELISA试剂盒测定PFC中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)水平;甲苯胺蓝染色观察PFC神经元形态;透射电镜观察线粒体微观结构和自噬情况;Western blot检测线粒体自噬相关蛋白PTEN诱导激酶1(PINK1)、parkin和微管相关蛋白1轻链3(LC3)的表达。结果:(1)与control组比较,aging组大鼠PFC中SOD和GSH-Px水平显著下调(P<0.01),MDA含量显著增加(P<0.01),Morris水迷宫定位航行实验中逃避潜伏期延长,穿越原平台的次数显著减少(P<0.01),证明衰老模型建成;aging组大鼠PFC神经元损伤显著,尼氏体数量减少,线粒体出现损伤;线粒体自噬相关蛋白PINK1和parkin表达受到抑制(P<0.05或P<0.01),LC3-II/LC3-I比值下降(P<0.01)。(2)与aging组比较,aging+Exe组大鼠PFC中SOD和GSH-Px水平显著性上调(P<0.01),MDA含量显著性降低(P<0.01),aging+Exe组大鼠逃避潜伏期缩短,穿越原平台的次数显著性增加(P<0.05),PFC神经元损伤减弱,尼氏体增多,线粒体结构改善,自噬体增多;线粒体自噬相关蛋白PINK1、parkin和LC3-II表达显著性上调(P<0.05,P<0.01)。结论:8周的间歇有氧运动可改善衰老模型大鼠空间学习记忆能力,可能与降低衰老大鼠PFC氧化应激,激活线粒体自噬相关蛋白PINK1、parkin和LC3-II的表达,改善线粒体结构和自噬水平有关。AIM To investigate the effect of aerobic interval exercise on mitophagy in the prefrontal cortex(PFC),and spatial learning and memory ability in a rat model of aging caused by D-galactose(D-Gal).METHODS Eight-week-old specific pathogen free(SPF)-grade Sprague-Dawley(SD)rats were randomly divided into control group,aging group,and aging+exercise(Exe)group,with 10 rats in each group.The rats in aging group and aging+Exe group received 6-week injection of D-Gal,while those in control group was injected with the same volume of saline.Subsequently,8 weeks of aerobic interval exercise intervention at 45 min/d for 5 d/week was performed in aging+Exe group.During this period,the rats in control group and aging group were naturally reared.After the exercise intervention,Morris water maze behavioral training and testing were performed.ELISA kits were used to determine the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in the PFC.Toluidine blue staining was used to observe the morphological changes of PFC neurons.Transmission electron microscopy was used to observe mitochondrial structure and autophagy.The expression of mitophagy-related proteins PTEN-induced kinase 1(PINK1),parkin and microtubule-associated protein 1 light chain 3(LC3)was determined by Western blot.RESULTS(1)Compared with control group,the levels of SOD and GSH-Px were significantly decreased(P<0.01),and MDA content was significantly increased(P<0.01)in the PFC of rats in aging group.The escape latency was prolonged and the number of crossings of the original platform was significantly reduced(P<0.01)in the Morris water maze positioning navigation experiment,demonstrating that the aging model was successfully generated.Neuronal damage in the PFC of rats in aging group was significant,the number of Nissl bodies was reduced and mitochondria showed damage.The expression of mitophagy-related proteins PINK1 and parkin was inhibited(P<0.05 or P<0.01),and the LC3-II/LC3-I ratio was decreased(P<0.01).(2)Compared with agin

关 键 词：线粒体自噬 学习 记忆 前额叶皮质 间歇有氧运动 衰老 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]