Research On The Effect And Mechanism Of Electrical Acupuncture On The Spatial Learning Ability And Synaptic Damage Of AD Rats Based On PI3K/Akt Signal Pathway

Objective:To observe the effects of electroacupuncture at Yin tang and Ying xiang on both sides of AD rats’spatial learning ability,hippocampal synaptic structure,and hippocampal phosphorylated PI3K/Akt protein expression level,aiming to further explore the effects of electroacupuncture on AD rats The biological mechanism provides basis and ideas for the treatment of clinical AD patients.method:1.Sixty-five adult male SD rats were selected.After the open field experiment,one unqualified rat was eliminated,and 12 were randomly selected as the normal group and 12as the sham operation group.The remaining 40 rats were modeled with Aβ_1-42,2 rats that died after modeling,and 2 rats that failed after modeling were eliminated,and the remaining 36 rats were divided into 3 groups:model group,Electroacupuncture group,medicine group,each group has 12 rats The sham operation group was prepared by injecting equal doses of normal saline into the hippocampus;the electroacupuncture group was used to stimulate Yin tang and bilateral Ying xiang points for intervention;the drug group was treated with donepezil hydrochloride at a dose of 3 mg/kg/d.Gavage;model group,sham operation group and normal group were given grasping stimulation at the same time and degree every day without any intervention.Course of treatment:For each group of rats,intervention was performed once a day,5 days as a course of treatment,2days intermittently,4 courses of treatment were observed.2.Experimental and index detection methods:the open field experiment was used to eliminate unqualified rats with anxiety;the Morris water maze experiment was used to screen the successfully prepared rats of dementia and the positioning navigation experiment to evaluate the changes in the learning ability of experimental rats;the hippocampal area was observed by transmission electron microscope The structural changes of synapses;immunohistochemical method was used to detect the expression of phosphorylated PI3K and Akt positive cells in the region of the rat hippocampus;the expression of PI3K and Akt protein in the region of the rat hippocampus was detected by Western blotting.result:1.The daily behavioral observation of rats shows that:Rats in the normal group and the sham-operated group have shiny hair,no significant changes in the amount of food and water during the experiment,free movement,and good reaction ability;the model group has dry and dull hair,significantly reduced eating and drinking,slow and awkward movements,The reaction ability was significantly reduced;the hair gloss of the electroacupuncture group and the drug group gradually improved,the food and water consumption gradually improved,and the action and reaction degree gradually increased.Situation and behavioral performance.2.The results of the water maze positioning and navigation experiment show:Compared with the blank group,compared with the blank group,the model group’s escape latency was significantly longer（P<0.01）;compared with the model group,the electroacupuncture group and drugs The escape latency of the group was significantly shortened（P<0.05）;this suggests that both electroacupuncture and donepezil hydrochloride intervention can improve the spatial learning ability of AD model rats.3.Synapse display of hippocampus under transmission electron microscope:The synaptic interface and bilayer membrane structure of the blank group and the sham-operated group were clear,the boundaries between the presynaptic and posterior membranes and the synaptic space were clear,and the vesicles were densely distributed.The synaptic membrane structure of the model group is incomplete,the synaptic interface fusion disappears,the boundaries between the presynaptic and posterior membranes and the synaptic gap are mostly blurred,and the vesicles in the presynaptic membrane are significantly reduced and unevenly distributed;the hippocampal synaptic membrane was partially dissolved,the structure was partially intact,the synaptic interface was clear,and the boundaries between the pre-synaptic and posterior membranes and the synaptic gap were improved to varying degrees and tended to become clear.Vesicles located in the presynaptic membrane gradually increase and tend to be densely distributed.This shows that electroacupuncture and donepezil hydrochloride can reshape the synaptic structure of the hippocampus of early dementia rats and play a role in improving the cognitive function of early dementia rats.4.Detect the expression of p-PI3K and p-Akt positive cells in the rat hippocampus by immunohistochemistry:Compared with the blank group,the expression of p-PI3K and p-Akt positive cells in the model group was significantly reduced（P<0.01）;Compared with the model group,the expression of p-PI3K and p-Akt positive cells increased in the electroacupuncture group and the drug group（P<0.05）;this indicates that the intervention of electroacupuncture and donepezil hydrochloride can increase the p-PI3K and p-Akt protein expression of AD model rats.5.Western blotting was used to detect the expression levels of p-PI3K and p-Akt proteins in the hippocampus of rats:Compared with the blank group,the expressions of p-PI3K and p-Akt proteins in the model group were significantly reduced（P<0.01）;compared with the model group In comparison,the expression of p-PI3K and p-Akt protein in the electroacupuncture group was significantly increased（P<0.05）,while the expression of p-PI3K and p-Akt in the drug group had no significant difference（P>0.05）;Acupuncture can increase the expression of p-PI3K and p-Akt protein in AD model rats.Conclusion:1.Through the intervention of electroacupuncture and western medicine donepezil hydrochloride,the behavioral performance,spatial learning ability,synaptic structure of hippocampus,and p-PI3K and p-Akt protein expressions of AD model rats prepared by injection of Aβ1-42 in the hippocampus were all Improved to varying degrees.2.The effect of electroacupuncture intervention in AD model rats is related to the PI3k/Akt signaling pathway,which may be through"increasing the expression of PI3K/Akt protein—promoting the repair of synaptic structure damage in the hippocampus—improving the spatial learning ability of AD model rats"The chain plays an effect.