| Objective:To explore the mechanism of electroacupuncture through mi RNA-132 to regulate the expression of downstream proteins Rac1 and Cdc42,enhance synaptic plasticity,improve the number and morphology of dendritic spines,and improve the ability of sleep deprivation of learning and memory in rats.Provide experimental basis for effective intervention targets of acupuncture in improving the decline of learning and memory ability caused by sleep deprivation,and provide new ideas and methods for enriching the role of acupuncture in the field of brain cognition.Methods:60 SPF male SD rats were randomly divided into 6 groups: blank group,model group,model + electroacupuncture group,model + sham electroacupuncture group,mi R-132-agomir electroacupuncture group,mi R-132-antagomir electroacupuncture group.In the blank group,the rats in this group were treated with the large platform water environment method without electroacupuncture treatment while the other groups were deprived of sleep;the model group was treated with the modified multiplatform water environment method to deprive the rats of this group without electroacupuncture.Treatment;model + electro-acupuncture group,using the modified multi-platform water environment method to sleep deprivation of this group of rats,sleep deprivation while giving electroacupuncture treatment;model + sham electro-acupuncture group,using the modified multi-platform water environment method to the rats Perform sleep deprivation and give false electroacupuncture intervention at the same time as sleep deprivation;mi R-132-agomir electroacupuncture group,use brain stereotaxic instrument to inject mi R-132-agomir into the hippocampus of rats for in vivo transfection,and then use the modified multi-platform It was sleep deprived by the water environment method and given electroacupuncture treatment;mi R-132-antagomir electroacupuncture group was injected with mi R-132-antagomir into the rat hippocampus using a stereotaxic apparatus to achieve transfection in vivo,and then the modified multi-platform was used The water environment law deprived him of sleep and was given electroacupuncture treatment.Rats started treatment on the first day of sleep deprivation.Rats in the electroacupuncture group were treated with electroacupuncture at "Baihui" and "Dazhui" at 9:00 a.m.for 15 minutes each time.5 days.In the sham electroacupuncture group,acupuncture points at "Dazhui" and "Baihui" acupuncture 3mm apart.The electroacupuncture instrument was also connected but not powered.The rest of the methods were the same as the electroacupuncture group.After 5 days of REM sleep deprivation and electroacupuncture treatment,Morris water maze positioning navigation experiment and space exploration experiment test were performed on the rats in each group to test the learning and memory ability of the rats in each group.After the last behavioral test,the rats were sampled,and then PCR fluorescence technology was used to detect the expression level of mi R-132 in the hippocampus of each group;Western blot was used to detect the protein expression of Rac1 and Cdc42;Golgi staining was used to detect the number of dendritic spines in hippocampal CA1 region of rats.Results:1.The general behavioral changes of rats after sleep deprivation.The rats in the blank group had normal food intake,red and translucent eyes,smooth and shiny hair,and free movement.Rats in each group using the improved multi-platform sleep deprivation had different degrees of food intake,eyes were dizzy,tired and fatigued,and their fur was rough and dull.2.The rat Morris water maze experiment results showed that: compared with the blank group,the model group had a longer average escape latency in the last test(P <0.01),and the number of crossing platforms and the target quadrant stay time were less(P <0.01,P<0.01);compared with the model group,the average escape latency of the rats in the model+ electroacupuncture group was shorter in the last test(P <0.01),and the number of crossing platforms and the target quadrant stay time were more(P <0.01,P <0.01);Compared with the model + electroacupuncture group,the average escape latency of the model + sham electroacupuncture group was longer in the last test(P<0.05),the number of crossing platforms and the target quadrant stay time were less(P<0.05,P<0.05),The mi R-132-agomir electro-acupuncture group had a shorter average escape latency in the final test(P<0.05),the number of crossing platforms and the target quadrant stay time was more(P<0.01,P<0.05),the mi R-132-antagomir electro-acupuncture group had the last The average escape latency of the test was longer(P<0.05),the number of crossing platforms and the stay time of the target quadrant were less(P<0.01,P<0.05).3.The results of PCR detection of mi R-132 expression showed that: compared with the blank group,the expression level of mi R-132 in the model group was lower(P<0.01);compared with the model group,the expression level of mi R-132 in the model +electroacupuncture group was higher High(P<0.01);Compared with the model +electroacupuncture group,the model + false electroacupuncture group and mi R-132-antagomir electroacupuncture group had lower mi R-132 expression levels(P<0.01,P<0.01),mi R-132-agomir electroacupuncture group mi R-The expression level of132 was higher(P<0.05).4.The relative expression of Rac1 and Cdc42 protein in each group of rats showed that compared with the blank group,the expression of Rac1 and Cdc42 in the model group was lower(P<0.01,P<0.01);Compared with the model group,the expression levels of Rac1 and Cdc42 in the model + electroacupuncture group were higher(P<0.05,P<0.01);Compared with the model + electroacupuncture group,the model + sham electroacupuncture group had lower expression levels of Rac1 and Cdc42(P<0.05,P<0.05),and the mi R-132-agomir electroacupuncture group had higher expression levels of Rac1 and Cdc42(P<0.05,P<0.05),mi R-132-antogomir electroacupuncture group had lower expression of Rac1 and Cdc42(P<0.05,P<0.01).5.The results of Golgi staining to detect the number of dendritic spines in CA1 area of hippocampus showed that compared with the blank group,the number of dendritic spines in the model group was smaller(P<0.01);compared with the model group,the number of dendritic spines in the model + electroacupuncture group was larger(P<0.05);Compared with the model + electroacupuncture group,the model + false electroacupuncture group and mi R-132-antagomir electroacupuncture group had fewer dendritic spines(P<0.05,P<0.05),mi R-132-agomir electroacupuncture group dendritic spine The number is large(P<0.05).Conclusion:1.The improved multi-platform water environment method can effectively deprive rats of sleep,which can increase the average escape latency,reduce the number of crossing platforms and target stay time,and significantly reduce learning and memory.2.Sleep deprivation can reduce the expression level of mi R-132 and reduce the activity of Rac1 and Cdc42 in rat hippocampus,thereby reducing synaptic plasticity.3.Electroacupuncture points "Baihui" and "Dazhui" can significantly improve the learning and memory abilities of rats with sleep deprivation,while fake electricity has little effect on the learning and memory abilities of rats.4.Overexpression of mi R-132 can increase the activity of Rac1 and Cdc42,enhance synaptic plasticity,and thus improve learning and memory.Inhibiting the expression of mi R-132 can reduce the activity of Rac1 and Cdc42,and decrease learning and memory.5.Electroacupuncture to improve the learning and memory ability of sleep deprived rats may be achieved by regulating the mi R-132-Rac1/Cdc42 pathway,increasing the number of dendritic spines,and regulating synaptic plasticity. |
