| The aging population has been expanding in the whole world, and this tendency is irreversible in decades. With aging the gradual emergence of cognitive function, especially the decline in the function of learning and memory, emotion regulation. The movement has neuroprotective effect on brain aging, but the mechanism is still far from clear. Autophagy is a creature in its development, aging exists in the process of a purifying their excess or damaged organelles common mechanism, life thereby to maintain protein homeostasis and cellular environment stable. This process in cell clear waste, structure reconstruction, growth development have played an important role. Studies have shown that exercise not only can promote autophagy in aging brain cells, the recession will also improve the ability of learning and memory of brain aging. Therefore, the movement to promote brain aging autophagy may be decreased the ability of learning and memory caused by the aging of the central nervous system, while in motion to improve aging brain learning plays a key role in the process of memory decline.The aim of study:Pay attention to the influence of motion reconstruction of brain aging morphology, metabolism and function of autophagy, in order to promote physical and mental health of the elderly, help the elderly and provide a theoretical basis for the realization of healthy aging and successful brain aging.Methods:Select 40 SD(Sprague Dawley) 18 month old healthy aged male rats were divided into aged control (OC) group, aging exercise (OS) group, aging exercise+saline ((OS+saline) group, aging exerciset chloroquine (OS+CQ) group, and another three month old SD rats as young control(YC) group.Exercise rats were swimming training for 8 weeks. All the rats were water maze training after the end of 48h. OS, OS+OS, saline+CQ group of rats for a period of 9 weeks of swimming exercise training. After the training, all the rats were trained to carry out water maze test, and the ability of learning and memory was detected by 48h. The expression of autophagy related protein LC3, p62, Atg5 and Atg7 were determined by Western blot.Western blot (Western blot) method for the determination of the expression of autophagy related protein LC3, p62, Atg5,Atg7.Results:1 Morris water maze showed that:(1)Comparison of the results of the escape latency time in the experiment of positioning navigation in the rats of each groupAfter 3 days of navigation experiments, rats in each group gradually reduced the latency period. In the water maze navigation experiment, compared with the YC group rats, OC group rats escape latency was significantly higher than that of the YC group rats (P<0.05); compared with the OC group rats, OS group, OS+saline group, OS+CQ group big mouse escape latency decreased significantly (P<0.01); compared with the OS+saline group, OS+CQ rats escape latency was significantly increased (P< 0.05).(2)Comparison of the results of the middle course of the navigation experiment in each groupAfter 3 days of directional navigation experiment, the rats in each group showed a downward trend. On the first day of the navigation experiment, there was no significant difference in the total distance of rats in each group. In navigation Experiment 2,3 days, compared with YC group, OC group, OS+CQ group rats in the navigation experiments in total distance increased significantly (P<0.01, P<0.01); compared with the OC group rats, the OS group, OS+saline group and OS+CQ group of rats were the total distance appears to have decreased significantly (P<0.01); compared with the OS+ saline group rats, OS+CQ rats total distance in the 3 days significantly increased (P< 0.01).(3) Comparison of the results of the target quadrant in space exploration experimentNavigation at the end of the experiment was performed 24 hours after the spatial probe test..The results showed that compared with the YC group, falling for the OC group and OS+CQ rats after the correct rate of the target quadrant (P<0.01); compared with the OC group rats, OS group, OS+saline group and OS+CQ rats after the correct rate of the target quadrant significantly increased (P<0.01); compared with the OS+saline rats. OS+CQ rats after the correct rate of the target quadrant significantly decreased (P<0.01).2 Western blot:(1)Detection of LC3 expression in hippocampus of rats by Western blot:Compared with the YC group rats, OC group and OS+CQ group rats hippocampus LC3Ⅱ/LC3I ratio was significantly higher than that of YC group, with a significant difference (P<0.01); Compared with the OC group rats, the ratio of OS group and OS+saline rats LC3-Ⅱ/LC3-Ⅰ was significantly lower than that of the OC group, with significant difference (P<0.01); Compared with the OS+saline group rats, the ratio of OS+CQ rats LC3-Ⅱ/LC3-Ⅰ was significantly higher than that of OS+saline group, with a significant difference (P<0.01).(2)Detection of P62 expression in hippocampus of rats by Western blot:Compared with the YC group rats, and the number of P62 in OC group, OS group, group OS+saline, OS+CQ group rats hippocampus significantly higher than YC group, with significant difference (P<0.01); Compared with the OC group rats, the number of P62 in OS group and OS+saline group rats hippocampus significantly lower than OC group, with significant difference (P<0.01); Compared with the OS rats, number of P62 in OS+CQ group in hippocampus of rats was significantly higher than that of OS+saline group, with a significant difference (P<0.01).(3)Detection of ATG5 expression in hippocampus of rats by Western blot: Compared with YC group rats, the expression of ATG5 in OC group, OS group, OS+saline group rats hippocampus significantly lower than YC group, with significant difference (P<0.01); Compared with the OC group rats, the expression of ATG5 in OS group, group OS+saline, OS+CQ group rats hippocampus significantly higher than OC group, with significant difference (P<0.01); Compared with the OS+saline rats, expression of ATG5 in OS+CQ group in hippocampus of rats was significantly higher than that of OS+saline group, with a significant difference (P<0.01).(4)Detection of ATG7 expression in hippocampus of rats by Western blot:Compared with the YC group rats, Atg7 in OC group, OS group, OS+saline group rat hippocampal expression appeared significantly decreased (P<0.01, P<0.05); Compared with the OC group rats, the expression of Atg7 in OS group, group OS+saline, OS+CQ group rats hippocampus appeared increased significantly (P<0.05, P<0.01); Compared with the OS+sasline group rats, the expression of Atg7 in OS+CQ rats hippocampus appeared significantly increased (P<0.01).3 immunohistochemical results(1) The expression of LC3 in hippocampus of rats in each groupCompared with YC group rats, OC, OS group, OS+saline group and OS+CQ group rats hippocampus LC3 positive autophagic particle aggregation number was significantly higher than that of the YC group; and OC group rats compared, compared to OS group, group OS+saline, OS+CQ group in hippocampus of rats LC3 positive autophagic particle aggregation number was significantly lower than that in the OC group rats; and group OS+saline rats. OS+CQ group in hippocampus of rats LC3 positive autophagic particles aggregation number was significantly lower than that of OS+saline rats. By means of CO staining with NeuN, the results of confocal microscopy showed that the expression of LC3 positive granules existed in rat neurons, and the presence of LC3 and NeuN were Co located.(2) The expression of P62 in hippocampus of rats in each groupA total of confocal microscopy observation showed that:compared to the p62 protein in the hippocampus showed punctate aggregations, compared with YC group rats, OC, OS group, OS+saline group and OS+CQ rats hippocampal p62 protein expression volume was significantly higher in the YC group; and the OC group rats compared to, the OS group, group OS+saline, OS+CQ rats hippocampal p62 protein expression number is significantly lower than that in the OC group rats; and group OS+saline rats. OS+CQ rats hippocampus p62 protein expression number were significantly lower than OS+saline rats. By CO staining with NeuN, the results of confocal microscopy showed that there was a co location of P62 and NeuN in rat neurons in the presence of P62.Conclusions:1 Long-term endurance exercise training improved the spatial learning and memory ability of aged rats.2 Autophagy inhibitor (Lysosome) partly abolished the effects of exercise on cognitive function in aged rats.3 Long-term endurance exercise training activates autophagy signaling in the hippocampus of aged rats, lysosome inhibitor partly abolished the effect of exercise.4 The improvement of the spatial learning and memory ability of aging rats may be related to the activation of autophagy. |
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