| ObjectiveAlzheimer’s disease(AD)is an age-related neurodegenerative disease characterized by progressive decline in memory and cognitive function,as well as behavioral and personality changes.A number of studies have shown that AD in the early stage has abnormal brain glucose metabolism before declining in cognitive function,which may be the key pathological changes of asymptomatic AD.Jinsiwei is a patent Chinese compound formula,which can tonify the Kidney,resolve Phlegm,invigorate Qi,and promote blood circulation,including ginseng,cistanche,rhizoma acori tatarinowii,curcuma aromatica and so on.Previous studies have shown that Jinsiwei can improve and delay the memory decline of AD patients,but the specific therapeutic target of AD in the early stage has not been studied.The aim of this study is to investigate the effect of the effect of traditional Chinese medicine prescription Jinsiwei on the early stage of AD mice by brain glucose metabolism,and to provide experimental basis for Jinsiwei treatment on early stage of AD.Materials and Methods1.Three-month-old APP/PS1 double transgenic mice were used as AD model mice,and they were randomly divided into 5 groups:the model group,the donepezil group,the Jinsiwei large,middle and low dose groups.C57/BL6J mice were used as the normal control group.The donepezil group mice were given donepezil hydrochloride(0.92mg/kg/d).The Jinsiwei large dose group mice were given Jinsiwei(20g/kg/d).The Jin siwei medium dose group mice were given Jinsiwei(10g/kg/d).The Jin SiWei low dose group mice were given Jinsiwei(5g/kg/d).The normal control group and the model group mice were given 0.5%CMC in the same volume.All the mice were given the drugs or CMC by intragastric administration once a day for 3 months.2.Morris water maze and platform were used to detect the behavioral changes of mice treated with JinSiWei after 3 months,and evaluate the effect of Jinsiwei on the spatial learning and memory ability of AD mice.3.The uptake of brain glucose in the temporal lobe,parietal lobe,frontal lobe and hippocampus of AD mice after 3 months of treatment was observed by Micro-PET.4.The changes of neurons and synaptic ultrastructures in hippocampal CA1 region of AD mice were observed by electron microscopy,which evaluated the effect of JinSiWei on the structures of neurons,synaptic and the amout of synaptic.5.The changes of GLUT1,GLUTS and PI3K,Akt,GSK3β and mTOR in AD mice were observed by immunohistochemistry,Western-blot and RT-PCR from the protein level and mRNA level that assessed effects of Jinsiwei on Glucose Transfection and Insulin Signal Transduction Pathway.Results1.The effect of Chinese Medicine Compound Jinsiwei on spatial learning and memory ability of AD miceIn the Morris water maze test,the escape latency of each group increased with training.The escape latency of the normal control group was significantly shorter than the model group(P<0.01).On the first and second day,compared with the model group,the treatment groups were no significant differences(P>0.05),but on the tired day,the escape period of the Jinsiwei low dose group was shorter(P<0.05).On the fourth and the fifth day,the treatment group was shorter or markedly shorter than the model group(P<0.01 or P<0.05).The swimming distance of each group in the search for hidden platform decreased with the increase of training times.Compared with the normal control group,the swimming distance of the model group were decreased from the first day to the fifth day(P<0.01 or P<0.05).On the first day and the second day,there was no significant difference between the treatment groups and the model group(P>0.05).On the third day,compared with the model group,the swimming distance of each dose group was decreased(P<0.01 or P<0.05).On the forth day and the fifth day,the treatment group were decreased or significantly decreased compared with the model group(P<0.01 or P<0.05).In space exploration experiment of the Morris water maze,the number of traversal platforms and the target quadrant residence time of model group in the 120s were significantly decreased compared with the normal control group(P<0.01).Compared with the model group,the target residence time of each treatment group were increased(P<0.01 or P<0.05).The number of crossing the platform of the middle and Jinsiwei low dose groups were increased compared to the model group(P<0.05).In the jump test,the number of errors of the model group was significantly increased compared with the normal control group(P<0.01).Compared with the model group,the number of errors of the donepezil group and the Jinsiwei middle dose group were decreased(P<0.05),and the number of errors of the Jinsiwei small dose group was significantly reduced(P<0.01).Compared with the normal control group,the latency time of the model group was significantly shorter.Compared with the model group,the latency time of the Jinsiwei group was prolonged or significantly prolonged(P<0.01 or P<0.05),and the donepezil group was no difference(P>0.05).2.The effects of Chinese herbal compound Jinsiwei on neurons and synaptic ultrastructure of AD miceElectron microscopic observated that the neurons and synaptic structures of normal control group mice in hippocampal CA1 were normal.The number of neurons in the hippocampal CA1 neurons of the model group was irregular,the electron density increased,and the chromatin margins were found.Golgi body was swelling,partial mitochondrial showed degeneration,lipofuscin significant increased,and the number of synapses in the nerve felt clearly decreased.Chinese medicine compound group could increase the number of synapses in the neural felt,and make protect with the neurons and synaptic structures.3.The effects of Chinese Herbal Medicine Jinsiwei on glucose uptake of AD mice Micro-PET was use for quantitative analysis of brain glucose uptake rate in the study area.the average brain glucose uptake rate of model group in four regions was decreased than the normal control group.Compared with the model group,the average brain glucose uptake in the middle and low dose group and the donepezil group was higher than that in the model group,but there was no significant difference between the high dose group and the model group4.The effects of Jinsiwei on glucose transport and Insulin Signal Transduction Pathway of AD miceWestern blot was used to detect GLUT1 protein expression in hippocampus.Compared with the model group,the expression of GLUT1 protein in the hippocampus of the normal control group was significantly increased(P<0.01).Compared with the model group,the expression of GLUT1 protein in the hippocampus of the donepezil group and the Jinsiwei middle dose group was increased(P<0.01),the Jinsiwei large dose and the Jinsiwei low dose group was increased(P<0.05).Immunohistochemical staining showed that the hippocampal CA1 positive cells were stained.The GLUT1 positive cells of the model group in the hippocampal CA1 region were significantly lower than those in the control group(P<0.01).Compared with the model group,the GLUT 1 positive cells in the treatment group were significantly increased(P<0.01).The expression of GLUT3 protein in hippocampus was detected by Western blot.The expression of GLUT3 protein of the normal control group in the hippocampus was significantly higher than the model group(P<0.01),and the expression of GLUT3 protein of the donepezil group,the Jin siwei large-dose and the middle-dose group was increased than the model group(P<0.05).Western blot was used to detect PI3K protein expression in hippocampus.Compared with the model group,the expression of PI3K protein in the hippocampus of the normal control group was significantly higher(P<0.05),and the expression of PI3K protein in the model group was s lower than that in the donepezil group(P<0.05).The expression of PI3K in the Jinsiwei middle dose and the high dose group was significantly increased than that in the model group(P<0.05 or P<0.01).Western blot was used to detect the expression of Akt protein in hippocampus.The results showed that the expression of Akt protein in the hippocampus of the model group was significantly lower than that in the normal control group(P<0.01).Compared with the model group,the expression of Akt protein in the hippocampus of the Jinsiwei high dose group was increased(P<0.05),and the expression of the donepezil group and the Jinsiwei middle-dose group significantly were increased(P<0.01).Immunohistochemical staining showed hippocampal CA1 positive reaction cells stained on cytoplasmic.the Akt-positive cells in the hippocampal CA1 region of the model group were significantly lower than those in the control group(P<0.01).Compared with the model group,the Akt positive cells in the treatment group were significantly increased(P<0.05 or P<0.01).The expression of p-GSK3β protein in hippocampus was detected by Western blot.The results showed that the protein expression of p-GSK3β in the hippocampus of the model group was significantly lower than that of the normal control group(P<0.01).Compared with the model group,the expression of p-GSK3β protein in hippocampus of the Jinsiwei hige dose and low dose group was increased(P<0.05),and the expression of p-GSK3βprotein in the donepezil group an the Jinsiwei medium dose group was significantly increased(P<0.01).Western blot was used to detect the expression of p-mTOR protein in hippocampus.The results showed that the expression of p-mTOR protein in the hippocampus of the model group was significantly higher than that of the normal control group(P<0.01).Compared with the model group,the expression of p-mTOR protein in hippocampus of donepezil group,the Jinsiwei high-dose and the low dose group was decreased(P<0.05),the medium-dose group was significant decreaed(P<0.01).The expression of PI3K mRNA and Akt mRNA was detected by RT-PCR.Compared with the model group,PI3K mRNA and Akt mRNA were higher expressed in the normal control group,and the expression of PI3K mRNA and Akt mRNA were also increased in the treatment group.ConclusionsChinese herbal compound Jinsiwei can improve glucose uptake rate of the specific brain area of,increase glucose transport,repair damaged insulin signaling pathway in the AD model mice,thereby improving brain glucose metabolism AD model mice in the early stage,and can maintain the structure neurons and synaptic,then improve the spatial learning and memory retention ability of the AD model mice in the early stage. |
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