The Effect Of Two Kinds Of Frequency Of The Electro-Nape-Acupunture On The Molecular Mechanism Of Space Learning And Memory Ability Of The Rats Model Of The Alzheimer’s Disease

Objective:To explore the effect of the molecular mechanism of the spatial learning and memory ability for the rats model of Alzheimer’s disease (AD)with two kinds of frequency of electro-nape-acupuncture, in order to better provide objective theoretical basis for the treatment of AD with this therapy method.Methods:20months of age, clean grade and male Sprague-Dawley (SD) rats feeding adaptation of one week, eliminateding the congenital dull rats by Morris water maze, were randomly divided into blank normal group, sham injury group, model group,2Hz and50Hz groups(n=10) based on body weight(320±50) from small to large numbers. Blank group rats were given a normal diet, drinking water. Sham group rats were given Normal saline with stomach feeding and intraperitoneal injection as the same with the model group. Model group and treatment group rats were given20mg/ml of AICl3solution2ml with stomach feeding and48mg/Kg of D-galactose with intraperitoneal injection every day, lasting8weeks, and feeding as same as blank group.After routine disinfection, take bilateral "Fengchi" and "Gongxue" as acupoints of the rats,with of Φ0.40X25mm acupuncture needles, piercing10mm, then connected with the pulse electro-acupuncture apparatus (DJ-6805D), ipsilateral upper and lower set of wires connected to the acupoints,which positive connected "Fengchi" and negative connected "Gongxue". The treatment group was divided according to the frequency2Hz and50Hz, energized stimulating contine20min, once a day, resting one day after6times. The therapy time was four weeks. Blank group, sham groupand model group were without treatment, but taking ahold and binding as the same time as the treatment group.Morris water maze test was used to the behavior changes of AD rats. HE staining and Electron microscope were used to look into the changes of histomorphology. Took Immunohisto chemistry and Insitu hybridization to detect the expression of pCREB1(mRNA), BDNF(mRNA), TrkB, Bcl-2/Bax, Caspase-3. The images were analyzed by the software of Motic image3.2, and the receipt were dealed with the SPSS17.0statistical software.Results:1. Behavior changes:Before modeling of rats in each group, the escape latency, the percentage of target quadrant swimming distance and the number of crossing the platform were difference (P>0.05). Compared with the blank and sham groups after the completion of the modeling, the escape latency, the percentage of target quadrant swimming distance and the number of crossing the platform of the model group,2Hz group and50Hz group were significant differences(P<0.01). After the end of treatment, compared with the blank and sham group, the behavioral indices of the model were significant differences (P<0.01); compared with the model group, the behavioral indices of2Hz group were difference(P<0.05), the50Hz group were significantly difference(P<0.01), compared with the2Hz group, the behavioral indices of50Hz group were difference (P<0.05).2. Tissue morphology:Compared with the blank group, the sham group changed little; the model group improved significantly. Compared with the model group, the degree of injury of the2Hz group were light, the50Hz group were lighter.3. Neuronal ultrastructure:there were no significant change between blank group and sham group. Lysosomes and lipofuscin of neurons increased, neuronal tigroid body, dispersion, mitochondrial cavitation, reduced synaptic vesicles.4The results of immunohistochemistry:(1) Compared with blank group and sham group, there were significant differences detected the positive cells expression of pCREB1, BDNF, TrkB, Bcl-2, Bax, Caspase-3protein of model group(P<0.01).(2) Compared with the model group, the positive cells expression of pCREB1, BDNF, TrkB, Bcl-2, Bax, Caspase-3protein of2Hz group were differences (P<0.05), and of50Hz group were significantly differences(P<0.01).(3) Compared with the2Hz group, the positive cells expression of pCREB1, BDNF, TrkB, Bcl-2, Bax, Caspase-3protein of50Hz group were differences (P<0.05).5The results of in situ hybridization:(1) Compared with control group and sham group, the positive cells expression of pCREB1mRNA and BDNFmRNA of model group had significant difference(P<0.01).(2) Compared with the model group, the positive cells expression of pCREB1mRNA and BDNFmRNA of2Hz group had difference (P<0.05), and50Hz group were significantly differences(P<0.01).(3) Compared with2Hz group, the positive cells expression of pCREBlmRNA and BDNFmRNA of50Hz group were differences (P<0.05).Conclusion:1. Two frequency of electro-nape-acupuncture can shorten the average escape latency, improved the times of crossing platform and the percentage of the target quadrant swimming path of the rats model of AD, and the effect of50Hz group was better than2Hz group.2. Two frequency of electro-nape-acupuncture can improve the morphology of hippocampal neural cells and ultrastructure of the rats model of AD, and the effect of50Hz group was better than2Hz group.3. Two frequency of electro-nape-acupuncture can increase the positive cells expression of the mRNA of pCREB1and BDNF, and of the protein of pCREB1, BDNF and TrkB of the rats model of AD, in order to improve the protection of hippocampal meuros, and the effect of50Hz group was better than2Hz group.4Two frequency of electro-nape-acupuncture can increase the positive cells expression of the protein of Bcl-2and reduce the positive cells expression of the protein of Bax and Caspase-3to reduce the apoptosis of hippocampal neurons, and the effect of50Hz group was better than2Hz group.5. The frequency’s speed of the electro-nape-acupuncture determined the therapeutic effect to the rats of AD, and the effect of50Hz group was better than2Hz group.