| Purpose:Research based on the theory of "Harmony between kidney and brain" cupping therapy for Alzheimer’s disease(Alzheimer ’s diseases,AD)model in mice hippocampal LC3-Ⅱ and cell autophagy related proteins in the mitochondria autophagy related protein Bnip3 effect,from the view point of mitochondrial autophagy "Harmony between kidney and brain" cupping therapy influence on "lack of brain" AD,explore its action mechanism,provide guidance and experimental basis for clinical treatment of AD.Material and method:The senescence acclerated mouse resistant(SAMR1)6 and the senescence acclerated mouse prone(SAMP8)18 were selected,and the SAMP8 mice were randomly divided into the model group,the western medicine group and the electroacupuncture group,each group of 6.Single cage feeding.1.Behavioral indicator detection: the Morris water maze was used to detect the learning ability and memory ability of mice,and it was divided into positioning navigation experiment and space exploration experiment.Positioning navigation experiment to avoid the incubation period and swimming total distance,to evaluate the learning ability of mice;The space exploration experiment measured the percentage of swimming time in the target quadrant,which was used to evaluate the memory capacity of the mice.2.Electroacupuncture treatment:control group,model group normal feeding,no treatment.In the western medicine group,the mice were injected with us dollars just for the stomach,and the mice were given the stomach one time per day.According to the theory of "Harmony between kidney and brain",the electroacupuncture group selected "baihui","shen shu" and "taixi" daily for 15 minutes.After the treatment,the therapeutic effect of electroacupuncture on AD model mice was evaluated by the change of behavioral indicators.3.After the index observation:behavioral index detection,anesthesia,put to death all the mice paraffin section of brain tissue in mice,using he staining and immunohistochemical,observed in mice hippocampal structure and nerve cell morphological changes;Separation ofhippocampus in mice,using Western Blot detection LC3-Ⅱ and Bnip3 content in the hippocampus.Results:1.After 8w of electric acupuncture and medication,Morris water maze navigation experiment results:with the increase of training days,groups of experimental mice to escape the incubation period are dwindling,the control group compared with model group,model group animal platform search efficiency drops,escape lengthen the incubation period,swimming distance increases,the difference was statistically significant.Including the western medicine group,compared with model group,western medicine group,the curative group animal platform search efficiency higher than that of model group,evade significantly shorten the incubation period,swimming distance than the model group decreased significantly,the difference was statistically significant.2.After 8w of electric acupuncture and medication,Morris water maze:space exploration results in the control group,western medicine group,the curative group target quadrant swimming time increases,the percentage of total time difference is statistically significant.3.After 8w of electric acupuncture and medication,Nepal’s dyeing test results:blank group of hippocampus CA1 area neurons of the vertebral body cell arrangement is compact,more layers of cells for 4-5 layer and the neuron cell body,pale cytoplasm and uniform,large,round nucleus,containing rich in austenite,darker.Model cones group of neurons in hippocampal CA1 area disordered arrangement,the number of chaos is not clear,the layer number of clutter,neuronal cell body shrinkage,missing or incomplete form,nissl bodies dissolved chaos.Western medicine group compared with model group,arrange relatively regular neurons,margin slightly loose,slightly fewer in number cell morphology,cell body shape size is normal,enriched with nissl bodies.The curative group compared with model group,neurons is orderly,slightly fewer in number cells form a slight shrinkage,form the cell body size is normal,enriched with nissl bodies,cytoplasmic staining and uniform,the large,round nucleus.4.After 8w of electric acupuncture and medication,immunohistochemical detection results:compared with control group,model group rats hippocampal CA1 area cell cytoplasm staining deep,is brown,the decrease in the number of cells,cell loosely arranged not neat,clearanceincreases,even a dropped cell layer,even some of the cell nucleus disappear,structure is not complete.Compared with model group,western medicine group,the curative group rats hippocampal CA1 area cell cytoplasm staining shallow,cells layer number,order,neuron structure is complete,cytoplasmic staining even and clear.5.After 8w of electric acupuncture and medication,Western Blot results:compared with model group,control group within the hippocampus LC3-Ⅱ way and Bnip3 protein content is lower than the model group(P<0.05);Compared with model group,western medicine,the curative group within the hippocampus LC3-Ⅱ and Bnip3 protein content is lower than the model group(P<0.05),the difference was statistically significant,the western medicine group compared with electric acupuncture group has no statistical significance.Conclusion:1.Under the guidance of the theory of "Harmony between kidney and brain",electroacupuncture therapy can effectively improve the learning and memory ability of AD model mice.2.Under the guidance of the theory of "Harmony between kidney and brain",electroacupuncture therapy is helpful for the normal operation of mitochondrial function,thus reducing the occurrence of autophagy in the mitochondria and making the cell morphology become normal.3.Under the guidance of the theory of kidney and brain of economic,cupping therapy can cut LC3-Ⅱ,the expression of Bnip3 protein content,lower cell autophagy and mitochondrial autophagy occurs. |
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