| Objective:Based on the previous PET-CT imaging results of the research group,acupuncture at the Dumai acupoint Baihui and Renzhong can improve glucose metabolism in the hippocampus of AD rats.Therefore,this experiment took AD rats as the research object,starting from the insulin PI3K/AKT signaling pathway,and explored the relevant mechanisms of acupuncture at the Dumai acupoint Baihui and humans in regulating glucose metabolism in the hippocampus of AD rats.Methods:80 SPF SD rats were selected and randomly divided into 4 groups: blank group,sham operation group,model group,and acupuncture group,with 20 rats in each group.D-galactose combined with Aβ 25-35 method was used for modeling.After successful modeling,the rats in the acupuncture group were acupunctured at Baihui and Shuigou once a day for 6 consecutive days and 1 day intermittent as a course of treatment,for a total of 6courses of treatment.The remaining three groups are only routinely crawled.The following indicators were observed after the acupuncture:(1)The Morris water maze test was used to evaluate the learning and memory ability of rats.(2)HE staining to observe the general structure and basic pathological changes of the hippocampus of each group of rats.(3)TUNEL staining to detect the apoptosis of nerve cells in the hippocampus of rats.(4)Immunohistochemical method was used to detect the expression of Aβ,Tau,p-tau,GLUT1,GLUT3,and GLUT4 protein in the hippocampus of rats.(5)Western blot method to detect the protein expression levels of insulin PI3K/AKT signaling pathway related index proteins INSR,PI3 K,AKT,GSK-3β in the hippocampus of rats.Results:(1)Behavioral test results:(1)In the positioning navigation experiment,compared with the blank group,the escape latency of rats in the model group was significantly longer(P<0.01);compared with the model group,the escape latency of rats in the acupuncture group was significantly shorter(P<0.01).(2)In the space exploration experiment,compared with the blank group,the number of times the rats in the model group crossed the platform was significantly reduced(P<0.01);compared with the model group,the times of the rats in the acupuncture group crossed the platform significantly increased(P<0.01).(2)The results of HE staining showed that the morphology of neurons in the hippocampus of rats in the blank group and the sham-operated group was normal,the nucleus structure was intact,and the cells were arranged in an orderly manner.No inflammatory cell infiltration,cell vacuole degeneration and cell necrosis were seen..In the model group,the neuron cells in the hippocampus of the model group are irregular,swollen,structurally disordered,and arranged unevenly,and vacuole-like cells appear.The neuron cell body shrinks,the nucleus shrinks,and the staining is light.Compared with the model group,the neuronal cell morphology of the acupuncture group was relatively normal,the structure was relatively complete,and the arrangement was neater.(3)TUNEL staining results showed that compared with the blank group,the apoptotic cells in the hippocampus of the model group increased significantly(P<0.01);compared with the model group,the apoptotic cells in the hippocampus of the acupuncture group significantly decreased(P<0.01)0.01).(4)Immunohistochemical staining results:(1)The expression of Aβ,Tau,and p-tau in the hippocampus of each group: Compared with the blank group,the expression of Aβ,Tau,and p-tau in the hippocampus of the model group was significantly increased(P<0.01);Compared with the model group,the expressions of Aβ,Tau and p-tau in the hippocampus of the acupuncture group were significantly reduced(P<0.01).(2)The expression results of glucose transporter related to the blood-brain barrier in the hippocampus of each group:Compared with the blank group,the expression of GLUT1,GLUT3 and GLUT4 in the hippocampus of the model group and the acupuncture group were significantly reduced(P<0.01);Compared with the model group,the expressions of GLUT1,GLUT3,and GLUT4 in the hippocampus of the acupuncture group were significantly increased(P<0.01).(5)Western blot test results: Compared with the blank group,the expression levels of INSR,PI3 K,and AKT proteins in the hippocampus of the model group were significantly reduced(P<0.01),and the expression levels of GSK-3β protein were significantly increased(P<0.01)Compared with the model group,the expression of INSR,PI3 K and AKT protein in the hippocampus of the acupuncture group was significantly higher(P<0.01),and the expression of GSK-3β protein was significantly reduced(P<0.01).Conclusion:(1)Acupuncture at Baihui and Shuigou can improve the learning and memory ability of AD rats.(2)Acupuncture at Baihui and Shuigou can alleviate the pathological damage of hippocampal neurons in AD rats and have certain brain protection effects.(3)Acupuncture at Baihui and Shuigou can repair and activate the central insulin PI3K/AKT signaling pathway,and effectively regulate the expression of key transporters of glucose metabolism in the hippocampus of AD rats,thereby comprehensively improving the level of glucose metabolism in the hippocampus of AD rats. |
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