| Objective:Alzheimer’s disease is a degenerative disease of the central nervous system,with progressive memory loss and cognitive dysfunction as the main clinical features,with insidious onset and long course of disease,which is a worldwide medical problem.In this study,guided by the theory of acupuncture and moxibustion for treatment of disease,with the neuroinflammatory response of AD as the starting point,the study and memory ability of AD-like model rats induced by D-galactose induced by pre-electricity was investigated from the perspective of cholinergic anti-inflammatory pathway The purpose is to clarify the mechanism of pre-electroacupuncture to improve the learning and memory ability and hippocampal neuroinflammatory response in AD-like rats,in order to provide experimental basis and theoretical support for the clinical promotion of acupuncture and moxibustion for prevention and treatment of AD.Methods:The experiment selected 36 4-month-old male SD rats and randomly divided them into a normal group,a model group,and a pre-electroacupuncture group,with 12 rats in each group.The rats in each group were raised in the same environment,and the normal group was not treated.The model group and the pre-electroacupuncture group were given D-galactose(150mg/kg/day)intraperitoneally for 8 weeks.In the pre-electroacupuncture group,daily intraperitoneal injection of D-galactose was followed by acupuncture at "Baihui" and bilateral "Zusanli" points.After acupuncture,the HANS-200 A electroacupuncture instrument was connected,and the electrode leads were connected to "Baihui" respectively.And “Zusanli” on one side,single-day connected to “Zusanli” on the left,Shuangri connected to “Zusanli” on the right,and “Zusanli” on the left and right sides alternately.The electroacupuncture instrument uses a continuous wave with a current of 1 m A and a frequency of 50 HZ.It is better to have rat limbs trembling and no hissing.The treatment is performed once a day,each treatment is 20 minutes,and the treatment is 6 days a week with one day off.Continuous treatment for 8 weeks.The normal group and model group are simultaneously grasped and fixed during the pre-electroacupuncture group treatment 20 min.After the intervention of each group,the Morris water maze experiment was used for behavioral testing.After the behavioral test,each group selected appropriate amount of rats for fresh tissue extraction and perfusion extraction.After sampling,the degree of activation of microglia in hippocampal CA3 area was observed by transmission electron microscope;the expression levels of inflammatory factors IL-1β,IL-6 and TNF-α in hippocampus were detected by ELISA method;microglia in hippocampus was detected by Western-Blot method The expression level of cell-specific protein Iba-1 and α7n ACh R;immunohistochemical method was used to detect the positive expression and localization of Ch AT in hippocampal CA3 area,and immunofluorescence was used to detect the expression of hippocampal microglia-specific protein Iba-1.Result:1.Pre-electroacupuncture can improve the learning and memory ability of AD-like model rats induced by D-galactoseMorris water maze experiment results: In the positioning navigation experiment,compared with the normal group,the escape latency of rats in the model group was significantly longer,and the difference was statistically significant(P<0.01);compared with the model group,the pre-electroacupuncture escape latency Significantly shortened,the difference was statistically significant(P<0.05).In the space exploration experiment,compared with the normal group,rats in the model group had a significantly reduced exploration time in the quadrant where the original platform was located,and the number of times they traversed the location of the original platform was significantly less,the difference was statistically significant(P<0.05);compared with the model group Compared with the electroacupuncture group,rats in the electroacupuncture group had significantly longer exploration time in the quadrant where the original platform was located,and the number of times they crossed the location of the original platform was significantly greater,and the difference was statistically significant(P<0.05).2.Pre-electroacupuncture can improve the neuroinflammatory response in the hippocampus of AD-like rats induced by D-galactoseELISA results: Compared with the normal group,the expression of inflammatory factors IL-1β,IL-6,and TNF-α in the hippocampus of the model group increased,and the difference was statistically significant(P<0.01);compared with the model group,the pre-electricity The expressions of inflammatory factors IL-1β,IL-6 and TNF-α in the hippocampus of the acupuncture group decreased,and the difference was statistically significant(P<0.05).3.Pre-electroacupuncture can inhibit the activation of microglia in the hippocampus of AD-like rats induced by D-galactoseImmunofluorescence results: Compared with the normal group,the number of activated microglia in the hippocampal CA3 area of the model group increased,and the fluorescence intensity increased,the difference was statistically significant(P<0.01);compared with the model group,pre-charge In the acupuncture group,the number of activated microglia in hippocampal CA3 area of rats in the acupuncture group decreased,and the average fluorescence intensity decreased.The difference was statistically significant(P<0.05).WB experiment results: Compared with the normal group,the relative expression of the specific protein Iba-1 activated by microglia in the hippocampus of the model group increased significantly,and the difference was statistically significant(P<0.01);compared with the model group The relative expression of Iba-1 in the hippocampus of the pre-electroacupuncture group was significantly reduced,and the difference was statistically significant(P<0.05).4.Pre-electroacupuncture can inhibit the activation of microglia in the hippocampus of AD-like rats induced by D-galactoseImmunohistochemical results: Compared with the normal group,the model group had fewer Ch AT positive cells in the hippocampus CA3 area,and the average optical density was lower(P<0.01);compared with the model group,the pre-electroacupuncture group had hippocampal CA3 Ch AT positive cells expressed more in the area,and the average optical density value was higher(P<0.01).WB experiment results: Compared with the normal group,the expression of α7 n ACh R protein in the hippocampus of the model group decreased,and the difference was statistically significant(P<0.01);compared with the model group,the expression of α7 n ACh R protein in the hippocampus of the pre-electroacupuncture group increased,The difference was statistically significant(P<0.05).Conclusion:1.Pre-electroacupuncture can improve the spatial learning and memory ability of AD-like rats induced by D-galactose.2.Pre-electroacupuncture can reduce the levels of pro-inflammatory factors IL-1β,IL-6 and TNF-α in the hippocampus of AD-like rats induced by D-galactose,indicating that pre-electroacupuncture can improve the nerves in the hippocampus of model rats Inflammatory response,which may be one of the mechanisms by which pre-electroacupuncture improves the learning and memory ability of AD-like rats.3.Pre-electroacupuncture can reduce the expression of specific protein Iba-1 which is activated by D-galactose-induced AD-like rats hippocampus microglia,indicating that pre-electroacupuncture can inhibit the small glue of AD-like rats The activation of plasma cells may be the mechanism by which pre-electroacupuncture improves the neuroinflammatory response in the hippocampus of rats.4.Pre-electroacupuncture can up-regulate the expression levels of cholinergic anti-inflammatory pathway related proteins Ch AT and α7n ACh R in hippocampus of AD-like rats induced by D-galactose,indicating that the mechanism of pre-electroacupuncture to improve the learning and memory ability of AD-like rats may be through activating the cholinergic anti-inflammatory pathway,inhibiting the activation of microglia in the hippocampus of rats,reducing the secretion of pro-inflammatory factors,and improving the hippocampus of rats Regional neuroinflammatory response,and then improve the learning and memory ability of rats. |
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