To Investigate The Effect Of Electricity On Autophagy Of Hippocampal Cells In Rats With Vascular Dementia Based On The PI3K/AKT/mTOR Pathway

Objective: Observation of the expression of proteins related to cognitive behaviors,pathological morphology of hippocampal CA1 area,hippocampal autophagy-related proteins Beclin1,LC3 and PI3 K / AKT / m TOR signaling pathway in rats with vascular dementia Impact,to explore the mechanism of electricity against VD.Methods: The VD rat’s model needed to make a duplicate of ischemia and reperfusion of the Common Carotid Arteries(CCA).The rats were randomly divided into the sham operation group and the waiting group,and the model group,the electroacupuncture 100 Hz group,the 2Hz group and the2/100 hz group.The electroacupuncture group needled "baihui","dazhui","geshu"(double)and "housanli"(double)points and connected the electroacupuncture apparatus.On the 17 th day after modeling,the behavior of rats was detected by the platform experiment.The pathological changes of hippocampal neurons were observed by HE staining,the Ultramicrostructure of hippocampus neurons was observed by transmission electron microscopy,and the expressions of Beclin1,LC3,PI3 K,p-AKT and p-m TOR proteins were detected by Western Blot.Results: 1.Jumping experiment: The learning and memory ability of rats in the model group decreased significantly(P<0.01).Electroacupuncture treatment group can improve the learning and memory ability of rats(P<0.01).The error frequency of rats in the electriroacupuncture 2Hz group and the 2/100 Hz group was less than that in the 100 Hz group,and the difference between the electroacupuncture 2/100 Hz group was significant(P<0.05,P<0.01).2.HE staining result : Neurons in the hippocampus of the sham group showed regular morphology,complete structure and orderly arrangement,without obvious shedding and loss.The cell morphology of model group was irregular,loose and disorderly,and shed obviously.The morphology of neurons in each electroacupuncture group was more regular and orderly,and the cells shed less.3.Transmission electron microscope results:In the sham-operated group,the nuclear membrane of hippocampal neurons was intact,and a small amount of cells were damaged.A large number of cells in the model group were swollen with mitochondria,and vacuoles and autophagosomes with double membrane structures were seen in neuronal cells.Mitochondrial swelling was reduced in each electiroacupuncture group,and a small number of neurons showed autophagosomes.4.Western Blot detection: The model group Beclin1 protein expression and LC3 Ⅱ/LC3 Ⅰ ratio are increased,PI3 K,p-AKT,p-m TOR protein expression were lower(P<0.01);The curative group can reduce Beclin1 protein expression and LC3 Ⅱ/LC3 Ⅰ ratio,increase PI3 K,p-AKT,p-m TOR protein expression(P<0.01).The curative group 2/100 Hz LC3 Ⅱ/LC3 Ⅰ below2 Hz and 100 Hz group(P<0.01),100 Hz PI3 K protein expression is higher than 2 Hz,the 2/100 Hz group(P<0.01),group 2 Hz,2/100 Hz p-AKT protein expression is higher than 100 Hz group(P<0.05,P<0.01),and 2 Hz,2/100 Hz group p-m TOR protein expression is higher than 100 Hz,2/100 Hz groups had significant difference(P<0.01).Conclusion: Electroacupuncture of "baihui","dahui","geshu" and "housanli" can improve the learning and memory ability of cerebral ischemia-reperfusion VD model rats and significantly improve the cognitive impairment of model rats.The mechanism may be to activate the PI3K/AKT/m TOR signaling pathway to inhibit the excessive activation of autophagy,so as to reduce the damage of hippocampal neurons and play a protective role in the brain.Moreover,the regulation of proteins related to autophagy and PI3K/AKT/m TOR signaling pathway and the improvement of cognitive function of model rats with different frequencies were different,and the frequency conversion and low frequency were better than high frequency.