Study On The Difference And Mechanism Of Different Acupuncture Intervention On AD Model Animal Based On "Enlightenment"

ObjectiveThis research aimed to observe the differences on 8-month SAMP8 mice between different kinds of acupuncture treatment. Based on this, explore the possible mechanism of "dredging DU meridian and awaking mind", especially in improving the inflammation of central nervous system.MethodSenescence-accelerated mouse prone 8 (SAMP8) and the cognate normal senescence-accelerated mouse-R1 (SAMR1) were housed under live conditions of controlled temperature (24 ± 2℃), a 12h/12 h dark/light cycle, and sterile drinking water and standard pellet diet ad libitum. Sixty 8-month-old SAMP8 male mice were divided into three groups (n=10 per group):SAMP8 Alzheimer’s disease control (AD) group, medicine control group (M), acupuncture group (A), medicine combined with acupuncture group (M&A), electro-acupuncture (EA) group and musical electro-acupuncture (MEA) group. Ten 8-month-old SAMR1 male mice were set as normal control (N) group. In the N group and AD group, do not do any treatment under the same rearing conditions, while grabbing and bondage the mice in order to ensure the same treatment conditions, once a day,15 days. In the medicine control group (M), the dosage of donepezil was 0.65μg/g per day. In the acupuncture group (A), acupuncture treatment was performed once a day for 15 days. The prescription of acupuncture points included DU20 Baihui, DU 26 Shuigou and EX-HN3 Yintang (the significant extra point). The needles were rotated at the rate of twice a second for 30s at each point. In the medicine combined with acupuncture group (M&A), the treatment was combined acupuncture treatment and donepezil treatment. In the EA group, the needle handle was connected with HANS-LH202 electro-acupuncture device, sparse wave,2 Hz of the frequency,2V of the voltage, and 0.6 mA of the current intensity. In the MEA group, the needle handle was connected with ZJ-12H Musical electro-acupuncture device. Rhythm clear, moderate speed and intensity of the music prescription (Curing dementia prescription) was selected, and adjusted the intensity so that the animals kept quiet during the course of treatment.After the treatment, Morris water maze (MWM) was used to test the spatial learning and memory ability. Micro-PET was used to test the glucose metabolism of the different regions of brain. HE staining was used to observe the changes in the dentate gyrus of hippocampus. Immunohistochemistry and Western blot method were used to analysis the NLRP3 inflammosome related protein (NLRP3, ASC, Caspase-1 and IL-1β).Results1.MWM:escape latency:compared with N group, from the first day to fifth day, other groups were higher than it (P<0.05). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.05). The times of crossing platform, ratio of swimming distance in the target quadrant, ratio of swimming time in the target quadrant:A group, M&A group, EA group and MEA group performed better than the AD group (P<0.05).2.Micro-PET:the ratio per gram of F-FDG in hippocampus:compared with N group, AD group and M group both lower than it (P<0.01). Compared with AD group, A group, M&A group, EA group and MEA group were higher than it (P<0.05). Compared with M group, M&A group, EA group and MEA group were higher than it (P<0.05). The ratio per gram of 18F-FDG in frontal lobe:compared with N group, AD group and M group both lower than it (P<0.01). The ratio per gram of 18F-FDG in parietal lobe:compared with N group, AD group and M group both lower than it (P<0.01).3.HE staining:N group:granular cell layer of granulose cells arranged in order, close, the cell body is oval, stained clear and uniform nuclei are round, clear nucleoli, basically no nucleolus shrinkage phenomena. AD group:cells scattered, irregular, cell shape is irregular, and mass cells nucleolus shrinkage, hyper chromatic cytoplasm; again, shape derangement of cell layers in the cells, nucleoli in some cells condensation phenomena.4.AP1-42 immunohistochemical analysis of integrated optical density (IOD) results: compared with N group, the IOD of AD group was higher than it (P<0.01). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). Compared with M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). Compared with A group, M&A group, EA group and MEA group were lower than it (P<0.01).5.NLRP3 inflammatory body-associated protein (NLRP3, ASC, Caspase-1, IL-1 Beta) Immunohistochemistry and Western blot analysis:NLRP3 relative expression levels: compared with N group, AD group was higher than it (P<0.01), M group, A group, M&A group, EA group and MEA group were higher than it (P<0.05). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). ASC relative expression levels:compared with N group, AD group was higher than it (P<0.01), M group, A group, M&A group, EA group and MEA group were higher than it (P<0.05). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). Caspase-1 relative expression levels:compared with N group, AD group was higher than it (P<0.01), M group, A group, M&A group, EA group and MEA group were higher than it (P<0.05). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). IL-1 beta relative expression levels:compared with N group, AD group was higher than it (P<0.01), M group, A group, M&A group, EA group and MEA group were higher than it (P<0.05). Compared with AD group, M group, A group, M&A group, EA group and MEA group were lower than it (P<0.01). Compared with M group, EA group and MEA group were lower than it (P<0.05).Conclusion1.Acupuncture intervention can improve spatial learning and memory abilities of dementia model mice. Effect of combination of acupuncture and medicine intervention is superior to a single drug (Donepezil) intervention. Effect of pulse electro-acupuncture and music electro-acupuncture is better than drugs, and music electro-acupuncture of the advantage is more obvious.2.Acupuncture intervention can improve the glucose metabolism in the hippocampus of dementia model mice. Effect of combination of acupuncture and medicine intervention is superior to a single drug (Donepezil) intervention. Effect of pulse electro-acupuncture and music electro-acupuncture is better than drugs, and music electro-acupuncture of the advantage is more obvious.3.Acupuncture intervention may improve Alzheimer’s mice neuron apoptosis in hippocampus dentate gyrus, it also can suppress the Aβ 1-42 expression and the effect was better than a single drug (Donepezil). Pulsed electro-acupuncture and music electro-acupuncture inhibition of aβ 1-42 Donepezil were better than both drugs and single intervention of acupuncture, and the advantage of music electro-acupuncture was more obvious.4. Acupuncture interventions can effectively inhibit expression of inflammatory factor IL-1 p in hippocampus of mice with Alzheimer’s. Effect of combination of acupuncture and medicine intervention is superior to a single drug (Donepezil) intervention. Effect of pulse electro-acupuncture and music electro-acupuncture is better than drugs, and music electro-acupuncture of the advantage is more obvious.5.The "dredging DU meridian and awaking mind" method can effectively inhibit the endogenous receptor of amyloid beta--NLRP3 inflammatory body and it’s associated protein (NLRP3, ASC, Caspase-1). And reduced the secretion of inflammatory cytokines IL-1 beta and release.