Protective Effect Of Huangqisan And Its Disassembled Prescriptions On Alzheimer's Disease Model Mice

Objective: In this study,5×FAD transgenic mice were used as the animal models to observe the effects of huangqisan and its disassembled prescriptions on learning and memory ability,metabolic indexes,?-amyloid plaques and the number of glial cells.Methods : 75 2-month old 5×FAD male mice and 15 wild-type C57BL/6J male mice born in the same litter were used as the normal control group.The 5×FAD mice were randomly divided into 5 groups with 15 for each group include huangqisan group,astragalus group,pueraria group,mulberry white group and model group.After 3 months of drug intervention,we started behavior test,including open-field experiment,new object recognition experiment and Morris water maze experiment to observe the changes of learning,memory and cognitive ability of mice.Then the metabolic cage was used to observe the change of respiration quotient,oxygen consumption,carbon dioxide production and activity.After that,we perfuse the mouse and take out its brain tissue to stain thio-s and observe the changes of ?-amyloid plaques area in mice.In addition,we take immunohistochemistry to observe the number of microglias and astrocytes in each group.Results: 1.Behavioral experiment results: The results of the open-field experiment showed that the moving distance and moving speed of the mice in pueraria group were higher than huangqisan group,with statistical significance(p<0.05).There was no significant difference in the percentage of marginal activity time.There was no difference between astragalus group,mulberry white group and huangqisan group.In addtion,there was no difference between the wild control group,model group and huangqisan group.Testing the new object recognition experiment at 1 hour,the identification index of astragalus group and puerarin group were obviously higher than model group(p<0.05).While the identification index of huangqisan group also has the rising trend,but no statistically significant.Testing the new object recognition experiment at 24 hour,we found that puerarin group was obviously higher than that the model group(p<0.05),while the identification index of huangaisan group and astragalus group also have the rising trend,but not statistically significant.No matter in which times,the white mulberry white group has no obvious change compared with other groups.There was no difference in the identification index between huangqisan and its splited prescription.Morris water maze experiment found that the escape latent period of mice in the huangqisan group,astragalus group and pueraria group were gradually shortened after the drug intervention.In addition,the percentage of target quadrant retention time in the huangqisan group and astragalus group were increased,which had statistical significance(p<0.05).Compared with the single drug,the huangqisan group showed an improvement trend,but no statistical significance.However,there was no statistical significance between the two group.There was no significant change in the mulberry white group compared with other groups.2.Various results of metabolic cage data test the wild group,5×FAD model group and huangqisan group showed that the oxygen consumption,carbon dioxide production,respiration quotient and activity level of mice in the evening increased compared with the daytime,which was consistent with the physiological activity characteristics of mice in the evening higher than in the daytime,but all of them were not statistically significant.3.The area ratio of ?-amyloid plaques in cortex area,hippocampal CA1 area and dentate gyrus areas in huangqisan group,astragalus group and pueraria group were significantly decreased compared with model group(p<0.05).Compared with astragalus group and pueraria group,huangqisan group had the best effect,but there was not statistically significant.The mulberry white group was statistically significant compared with the huangaisan group(p<0.05),but there was no significant difference between the mulberry white group and the model group.4.Cytopathological studies showed that Huangqisan group,astragalus group and pueraria group can reduce the proliferation and activation of microglias and astrocytes in the brain of 5×FAD transgenic mice.The results of microglia quantization showed that huangqisan had the best effect compared with astragalus group and pueraria group,but there was no statistical difference.But there was no significant difference between the mulberry white group and the model group.The results of astrocytes quantization showed that huangqisan and its disassembled prescriptions group had statistical significance compared with the model group(p<0.05).Neither microglia nor astrocytes was no significant difference between the model group and the mulberry white group.Conclusions: We found that the main drugs in huangqisan prescription for prevention and treatment of Alzheimer's disease model mice were astragalus and pueraria by taking the improvement degree of learning and memory ability and cognitive impairment of 5×FAD transgenic mice,the area occupied by ?-amyloid plaques and the number of glial cells as the observation standard.However,the mulberry white showed less effect on 5×FAD transgenic mice.