| Objective:1.To observe the improvement effect of electroacupuncture and gastrodin on the behavior of AD rats.2.To observe the effect of electroacupuncture on the expression of galanin receptor 2(GalR2)and acetylcholinesterase(AchE)in hippocampus and amygdala of AD rats.;To observe the effect of gastrodin on the expression of GalR2 and AchE in hippocampus and amygdala of AD rats.3.To explore the role of GalR2 and AchE in the treatment of AD by electroacupuncture and gastrodin,and the relationship between them,so as to provide basic theoretical basis for clinical treatment of AD.Methods:Forty adult male SD rats,weight 200-220g,were randomly divided into 8 groups with 5 rats in each group:normal group(group N),model group(group M),gastrodin+M871 group(group Gas+M871),gastrodin group(group Gas),gastrodin+M1145 group(group Gas+M1145),electroacupuncture+M871 group(group EA+M871),electroacupuncture group(group EA)and electroacupuncture+M1145 group(group EA+M1145).Group N did not receive any treatment.In group M,Aβ142 was microinjected into bilateral hippocampus by stereotactic device,5μl/side.In Gas group,gastrodin was injected intraperitoneally at a dose of 0.1ml/100g qd for 4 weeks.In Gas+M871 group and Gas+M1145 group,M871 and M1145 were injected into unilateral lateral ventricle at a dose of 5μl,once every 10 days,twice in total,and the other steps were the same as those in Gas group.EA group was given EA at "Dazhui","Baihui" and "Zusanli" with frequency of 2Hz,current intensity of 1mA,density wave,lasting for 30min,qd.EA+M871 group and EA+M1145 group were given unilateral intracerebroventricular injection of M871 and M1145,5μl,once every 10 days,a total of two times,the other steps were the same as EA group.The spatial and learning and memory abilities of rats in each group were observed and compared by water maze test.Nissl staining and immunohistochemical staining were used to observe the morphological and quantitative changes of GalR2 and AchE positive neurons in hippocampus and amygdala.Western blotting was used to detect the expression of GalR2 and AchE in hippocampus and amygdala.Results:1.The results of water maze test showed that the average escape latency of rats in group M was significantly prolonged(P<0.05),and the percentage of residence time in target quadrant was significantly shortened(P<0.05);the average escape latency of rats in each treatment group was shortened(P<0.05),and the percentage of residence time in target quadrant was increased(P<0.05),in which Gas+M1145 group and EA+M1145 group were the most significant,while Gas+M871 group and EA+M871 group were the most significant,the escape latency of was longer than that of Gas group and EA group(P<0.05),and the percentage of residence time in target quadrant was shorter(P<0.05).2.Nissl staining showed that the number and processes of neurons in hippocampus and amygdala of group M were decreased,and the cell bodies were vacuolated.The number of neurons in each treatment group was increased(P<0.05),and the cell morphology was improved.The number of cells in Gas+M871 group,Gas group and Gas+M1145 group showed an increasing trend(P<0.05);the number of cells in EA+M871 group,EA group and EA+M1145 group also showed an increasing trend(P<0.05).3.The results of immunohistochemistry showed that the number of GalR2 immunoreactive neurons in hippocampus and amygdala of group M was increased(P<0.05),and the number of GalR2 immunoreactive neurons in hippocampus and amygdala of each treatment group was further increased(P<0.05),especially in Gas+M1145 group(P<0.05);the number of AchE immunoreactive neurons in hippocampus and amygdala of group M was significantly increased(P<0.05)The number of cells decreased(P<0.05),especially in Gas+M871 group and EA+M871 group(P<0.05).4.Western blotting results showed that the expression of GalR2 protein in hippocampus and amygdala of rats in group M was increased(P<0.05),but there was no significant change in Gas+M871 group and EA+M871 group compared with group M(P>0.05),and the expression of GalR2 protein in other four groups was further up-regulated(P<0.05);the expression of AchE protein in hippocampus and amygdala of rats in group M was significantly increased(P<0.05)The expression of ache protein in each treatment group was significantly decreased(P<0.05).Conclusion:1.EA and gastrodin can improve the behavioral changes of AD model rats.2.Electroacupuncture and gastrodin could up regulate the expression of GalR2 in hippocampus and amygdala,and decrease the activity of ache.3.The interaction between GalR2 and AchE may affect the therapeutic effect of AD. |
