The Mechanism Study Of Electroacupuncture Activating AMPK To Affect Aerobic Glycolysis On Learning-memory Ability In APP/PS1 Mice

Objective:The study aimed to explore the mechanism of electroacupuncture?EA?at the Baihui and Shenting acupoints activating brain AMPK to regulate aerobic glycolysis?Warburg effect?of hippocampus,which enhanced glucose metabolism to improve the learning-memory ability in early Alzheimer's disease models of APP/PS1 transgenic mice.Methods:Breeding of 4-month-old SPF APP/PS1 transgenic mice,using PCR to identify the mice genotype.According to the PCR results,transgenic mice were divided into the APP/PS1group,the APP/PS1+AMPK^+/-group,the APP/PS1+EA group,the APP/PS1+AMPK^+/-+EA group,with 10 mice in each group,another 10 negative mice for wildtype?WT?group.Morris water maze and new object behavior were used to test the learning-memory ability of each group.In the APP/PS1+EA group and the APP/PS1+AMPK^+/-+EA group,EA at Baihui and Shenting acupoints were adopted with the frequency of 1/20hz,once a day for 30 minutes,5days a week for 4 weeks.The WT group,the APP/PS1 group and the APP/PS1+AMPK^+/-group were only grabbed under the same conditions,without any special treatment.Before and after the intervention,chemical exchange saturation transfer?CEST?imaging was used to observe the changes of glucose metabolism in bilateral hippocampal brain regions of each group.Western Bloting?WB?were used to observe the expression levels of AMPK and key enzymes of Warburg effect in the hippocampus of APP/PS1 transgenic mice.Results:?1?The effects of EA on the learning-memory of APP/PS1 transgenic mice.Before the intervention,new object recognition showed that compared with WT group,the learning-memory ability of mice in APP/PS1 group and APP/PS1+EA group decreased?P=0.034,P<0.01?,and there was no statistical difference between the two groups?P>0.05?.Compared with APP/PS1 group,the learning-memory ability of the APP/PS1+AMPK^+/-group and the APP/PS1+AMPK^+/-+EA group was worse?P=0.030,P=0.031?,and there was no statistical difference between the two groups?P>0.05?.After intervention,the water maze test showed that compared with WT group,the average escape latency of APP/PS1 group was significantly prolonged?P<0.01?,the time proportion and times of crossing the quadrant of the platform were less?P<0.01,P<0.01?.Compared with APP/PS1 group,the average escape latency of the APP/PS1+AMPK^+/-group was longer?P<0.05?,and the time proportion and times of crossing the quadrant of the platform were less?P=0.007,P=0.023?.However,compared with the APP/PS1 group,the escape latency time of APP/PS1+EA group was shorter?P<0.01?,and the time proportion and times of crossing the quadrant of the platform were increased?P=0.030,P=0.001?.Compared with the APP/PS1+AMPK^+/-group,the escape latency period of the APP/PS1+AMPK^+/-+EA group was significantly shorter?P<0.01?,and the time proportion and times of crossing the quadrant of the platform were increased?P<0.01,P<0.01?.Moreover,compared with the APP/PS1+AMPK^+/-+EA group,the escape latency period of the APP/PS1+EA group was significantly shorter?P=0.027?,and the times of crossing the platform was increased?P=0.039?.The above results suggested that the learning-memory ability of APP/PS1,APP/PS1+AMPK^+/-mice were decreased,and EA at Baihui and Shenting acupoints could improve learning-memory ability of APP/PS1 transgenic mice.?2?The effects of EA on hippocampal glucose metabolism of APP/PS1 transgenic mice.Before and after the intervention,MRI-CEST imaging was conducted.Before the intervention,there was no statistical difference in left and right glucose metabolism levels between the WT group,the APP/PS1 group,the APP/PS1+EA group,the APP/PS1+AMPK^+/-group,and the APP/PS1+AMPK^+/-+EA group?P>0.05?.After intervention,glucose metabolism in the left hippocampus of the APP/PS1 group,the APP/PS1+AMPK^+/-group were decreased compared with the WT group?P=0.001,P=0.001?.Compared with APP/PS1 group,the glucose metabolism levels of the APP/PS1+EA group in the left and right hippocampus were significantly increased?P<0.01,P<0.01?.Compared with APP/PS1+AMPK^+/-group,the APP/PS1+AMPK^+/-+EA group increased?P=0.026,P=0.030?.Compared with the APP/PS1+AMPK^+/-+EA group,the glucose metabolism levels of the APP/PS1+AMPK+EA group in the left and right hippocampus increased?P=0.012,P=0.037?.It suggestes that EA could improve the hippocampal glucose metabolism of APP/PS1 transgenic and AMPK heterozygous knockout mice.?3?The effects of EA on hippocampal AMPK expression and Warburg effect in APP/PS1 transgenic mice.Western Bloting results showed that compared with WT group,the expression levels of AMPK,PFKFB3,HK2 and PKM2 in APP/PS1 group were significantly reduced?P<0.05?.Compared with the APP/PS1group,the expression levels of AMPK,PFKFB3,HK2 and PKM2 were decreased in the APP/PS1+AMPK^+/-group?P<0.05?.Compared with the APP/PS1 group,the expression levels of AMPK,PFKFB3,HK2 and PKM2 in the APP/PS1+EA group were increased?P<0.05?.Compared with the APP/PS1+AMPK^+/-group,the expression levels of AMPK,PFKFB3,HK2 and PKM2 in the APP/PS1+AMPK^+/-+EA group increased?P<0.05?.Compared with the APP/PS1+AMPK^+/-+EA group,the expression levels of AMPK,PFKFB3,HK2 and PKM2 were increased in the APP/PS1+AMPK+EA group?P<0.05?.The above results showed that the AMPK activity of APP/PS1,APP/PS1+AMPK^+/-mice decreased in hippocampal brain region,and the expression of key protease of Warburg effect decreased in glucose metabolism.EA at Baihu and Shenting acupoints could activate AMPK to improve the expression levels of PFKFB3,HK2 and PKM2 in APP/PS1 transgenic mice.Conclusion:EA at Baihui and Shenting acupoints could improve the learning-memory ability by activating hippoacupal AMPK to regulate the key molecules of Warburg effect and improve glucose metabolism in APP/PS1 transgenic mice.