| Objective:In this study,we observed the effects of Electro-acupuncture on spatial memory ability and neuron autophagy level,neuron apoptosis ratio and Syn protein expression in hippocampal CA1 area of Rats with Alzheimer’s Disease,and to explore the possible mechanism of Electro-acupuncture on spatial memory ability of Rats with Alzheimer’s Disease,and to provide evidence for further study of electro-acupuncture for Rats with Alzheimer’s Disease.Method:1.After the open field trial,an unqualified rats were removed from 65 adult SD rats,and 24 rats were randomly selected from 64 qualified rats.They were randomly divided into normal group and sham operation group,12 rats in each group.The remaining 40 rats were injected Aβ1-42 into the lateral ventricle by reference to the literature to establish the model of Rats with Alzheimer’s Disease.The Morris water maze test was used to evaluate whether the model was successfully established,and 2 unqualified and 2 dead rats were excluded.The remaining 36 rats were randomly divided into three groups,namely,the model group,the EA group and the drug group,12 in each group.The sham operation group was injected the same dose of normal saline with the same method.After first days,the EA group was given electro-acupuncture to stimulate both sides of Ying Xiang and Yin Tang.The drug group was given gastric lavage with donepezil,while the normal group,the sham operation group and the model group did not receive any treatment.Only in the EA group,during the treatment,EA group was used for grasping and stimulating;the above groups were treated once a day,continuous treatment of 5 day for a courses,after 2day rest,start another treating,a total of 4 courses.2.After the treatment,The Rats in each group were trained with 3 day Morris water maze.Then the Morris water maze system was used to carry out the 3 day space exploration experiment to evaluate the memory ability of each group.3.After the last behavioral test,the samples were fixed and stained with Nissl.The Nissl bodies were observed under microscope to evaluate the expression and morphology of Nissl bodies.The autophagy level of neurons was evaluated.The percentage of neuronal apoptosis was observed by TUNEL staining,and the apoptosis level of neurons wasevaluated.The expression of Syn was observed by Western blot and immunohistochemistry,and the changes of synaptic plasticity were evaluated.Results:1.General situation comparison:after modeling,the normal group and the sham operation group of rats had smooth skin,normal food intake,and sensitive in response to stimulate;compared with sham operation group,the rats in the model group,EA group and drug group were scattered and rough,slow in action,and obviously retarded in the stimulation reaction.After intervening,the normal group and the sham operation group of rats had smooth skin,normal food intake,and sensitive in response to stimulate.Compared with the normal group and sham operation group,the rats in the model group were coarse,slow,and slow in response to stimulate.Compared with the model group,the skin of the EA group and the drug group were basically smooth and only a little scattered.The food intake was normal,and the response to stimulate was rapid.Compared with the EA group,there was no significant difference between the two groups in skin glossiness,food intake,and response to stimulate and so on.2.Behavioral examination results: after modeling,the normal group and the sham operation group: there was no significant difference in the time between the first crossing platform and the number of platforms across the platform in the two groups(p>0.05);compared with the sham operation group,the number of crossing platform and the time for the first crossing platform significantly increased in the model group,EA group and drug group(p<0.01).After intervening,the normal group and the sham operation group of rats had the time of the first crossing platform and the number of times across the platform(p>0.05);compared with the normal group and the sham operation group,the number of crossing platform in the model group decreased significantly,and the time for the first crossing platform increased significantly(p< 0.01);compared with the model group,the number of crossing platform in the EA group and the drug group increased significantly.The time for the first time across the platform was significantly reduced(p<0.05).Compared with the EA group,there was no significant difference in the time between the two groups of rats crossing the platform for the first time and the number of times across the platform(p>0.05).3.Nissl staining results:the number of Nissl bodies positive cells was not significantly different between the normal group and the sham operation group(p>0.05).Compared with the normal group and sham operation group,the number of Nissl bodies cells in the model group was significantly reduced(p<0.01).Compared with the model group,the number of Nissl bodies cells in the EA group and the drug group increased significantly(p<0.05).Compared with the EA group,there was no significant difference between thetwo groups in the number of Nissl bodies(p>0.05).4.TUNEL staining results: there was no significant difference in the proportion of neuron apoptosis between the sham operation group and the normal group(p>0.05).Compared with the normal group and sham operation group,the proportion of apoptotic neurons in the model group increased significantly(p<0.01).Compared with the model group,the proportion of neuron apoptosis in EA group and drug group decreased significantly(p<0.05).Compared with EA group,there was no significant difference between the two groups in the proportion of neuron apoptosis(p>0.05).5.Western blot and immunohistochemistry results: there was no significant difference in the relative expression of Syn protein and the number of positive expression between the normal group and the sham operation group(p>0.05).Compared with normal group and sham operation group,the relative expression quantity and positive expression quantity of Syn protein in model group were significantly decreased(p<0.01).Compared with model group,the relative expression quantity and positive number of Syn protein in EA group increased significantly(p<0.01).In the drug group,the relative expression quantity of Syn protein and the number of positive expression increased significantly(p<0.05).Compared with the EA group,the relative expression and positive expression of Syn protein in the drug group decreased significantly(p<0.05).Conclusion:1.The decrease of spatial memory ability in Rats with Alzheimer’s Disease is closely related to the decrease of autophagy level in hippocampus neurons,the increase of neuronal apoptosis ratio and the decrease of Syn protein expression.2.Electro-acupuncture and donepezil can regulate the autophagy level of hippocampal neurons in Rats with Alzheimer’s Disease,antagonizing the apoptosis of neurons induced by Aβ1-42 and increasing the expression of Syn,which may be one of the potential mechanisms of improving spatial memory impairment in Rats with Alzheimer’s Disease,but the mechanisms of action of the two may differ. |
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