Effect Of Electroporation On Behavior And Cortical Klotho Gene Expression In APP/PS1 Transgenic Mice

ObjectiveThe purpose of this study was to investigate the effects of the intervention methods of Baihui,Yin-Tang and Shuigou acupoints on APP-PS1 double transgenic mice at 6 months of age in order to provide new clinical treatment and the objective scientific basis for Alzheimer’s disease;and on this basis,explore the electric acupuncture on the 6-month-old APP/PS1 double transgenic mice at Baihui,Yintang,Shuigou acupuncture points to study whether by increasing Klotho gene expression to improve the learning and memory ability of Alzheimer’s disease(AD)model and to explore the mechanism of electroacupuncture treatment of Alzheimer’s disease.MethodsA total of 20 APP/PS1 transgenic mice aged 6 months were randomly divided into model group,electroacupuncture group and 10 C57BL/6 wild mice as normal control group.EA group included Baihui and Yintang in which Baihui tip back to the head and Yintang to the tip of the nose needling 2-3 mm.The use of density wave,the frequency of 1/50 Hz,intensity 0.3 mA,sustained 20 min;needleing handle tremor to the degree where animals do not struggle and no neighing for.At the end of electroacupuncture group,the point of Shuigou acupuncture points,with the normal group and the model group the same method with the mouse bag tied 15 min,every other day for 4 weeks.The Morris water maze was selected to study the spatial learning and memory abilities of mice in each group.The fourth quadrant of the sailing experiment was located at the escape point of the water landing point,the number of swimming through the platform and the third quadrant swimming distance were used to evaluate the ability index of learning and memory of mice.The expression of Klotho was observed by immunohistochemistry and the expression ability of Klotho was detected by RT-PCR.And use SPSS20.0 statistical software for statistical analysis.Results1.Morris water maze testMorris water maze test showed that the escape latency time of mice in each group was shortened(P<0.05),with the increase of training days;compared with the control group,the escape latency of the model group was prolonged and that of the electroacupuncture group was shorter than that of the model group.The number of crossing platform and the swimming distance of platform quadrant in each group were analyzed by ANOVA.Compared with the control group,the number of crossing platform and the distance of swimming in platform quadrant in the model group were significantly reduced.The number of crossing platform increased in electroacupuncture group(P<0.05).Compared with the normal control group,the escape latency increased in the model group,the times of the space exploration experiment crossing the platform,the swimming distance in the platform quadrant decreased significantly(P<0.05,P<0.01)and the escape latency in the electroacupuncture treatment group all decreased significantly(P<0.05).2.Effect of electroacupuncture on the expression of Klotho protein in APP/PS1 mice.In normal group,brown positive expression was found in Klotho cells,and in model group,the expression of Klotho was significantly lower than that in normal group,which only in the cells,compared with the model group,the positive expression of Klotho in the model group was higher than that in the control group.The expression of Klotho in the electroacupuncture group was significantly enhanced.The results showed that the expression of Klotho in the model group was lower than that in the normal group(P<0.01),but the expressionability of Klotho in the electroacupuncture group was higher than that in the model group(P<0.01).3.Effect of electroacupuncture on Klotho mRNA expression in APP/PS1 miceCompared with the control group,the relative expression of Klotho mRNA in the model group was significantly lower than that in the model group(P<0.01),and the relative expression of Klotho mRNA in the electroacupuncture group was higher than that in the model group(P<0.01).Electroacupuncture promoted the expression of Klotho mRNA in cerebral cortex.ConclusionEA Baihui,Yintang,Shuigou can improve the ability of learning and memory in APP/PS1 transgenic mice.Electroacupuncture treatment of APP/PS1 double transgenic mice can enhance the expression of anti-aging gene Klotho.The mechanism of which may promote learning and memory by regulating GluN2B,promoting NMDA receptor to open and receive information for a long time,and then maintaining the information exchange between cells so as to promote learning and memory.and maybe have a benign regulatory effect on the prevention and treatment of Alzheimer’s disease.