| Alzheimer’s Disease(AD)is an insidious and progressive neurodegenerative disease.Clinically,memory impairment,aphasia,deaconsia,dementia,impairment of visual skills,executive dysfunction,and generalized dementia such as personality and behavioral changes are characterized.Cognitive dysfunction(CD)is the beginning of progressive deterioration of AD.The treatment of CD is of great significance for the prevention and treatment of AD.Pathogenesis of CD is complicated,including kidney deficiency and hyperactivity of heart fire,heart and kidney disconnection.Jiao-tai-wan(JTW)is from the Ming Dynasty,"Han’s Medical Link" is a prescription for the treatment of insomnia,from Coptis,cinnamon two flavors of drugs,which with traffic heart and kidney effect.Coptis has the protection of neurons,improve nerve inflammation,relieve cognitive impairment caused by diabetes,cinnamon also has the protection of the central nervous system,improve the cholinergic system and other effects.Objective:We envisage that the JTW can protect the nervous system and improve the symptoms of cognitive dysfunction.Based on the above assumptions we conduct this experiment to improve efficacy of scopolamine model mice cognitive dysfunction and mechanism research.The study included behavioral studies to observe the effects of low,medium and high doses of JTW on the learning and memory of mice.Through the study of the effects of JTW on the brain tissue cholinergic system,oxidative stress system,related neuron cells and apoptosis to investigate the mechanism of improving the cognitive dysfunction of scopolamine model mice.Methods:96 male and female kunming mice were male and female,and their weight was 21-23 g,randomly divided into 6 groups.(1)using Morris water maze,Y maze,new object recognition,and conditional fear experiment,which to observe the changes of learning and memory function in mice.(2)using spectrophotometry to detect brain acetylcholine(Ach)levels in mice,acetylcholinesterase(AChE)and acetylcholine transferase(ChAT)activity and malondialdehyde(MDA)and reactive oxygen species(ROS)level,the total superoxide dismutase(SOD)and catalase(CAT)activity.(3)the expression of neurons PSD95,SYN,BDNF and NGF protein,as well as the expression of apoptotic protein bcl-2,anti-Bax and anti-Caspase-3 protein were detected by western blotting.(4)the changes of neurons and apoptosis were observed by Nissl and Tunel staining.Results:The main eight components of the preparation are Magnoflorine,Groenlandicine,Coptisine,Jatrorrhizine,Columbamine,Berberine,Palmatine,and Cinnamaldehyde.Among them,berberine and cinnamaldehyde were the effective components of Coptis chinensis and Cortex Cinnamomi,and berberine accounted for the highest proportion.In the escape incubation experiment,after the model group was given scopolamine,the cognitive function of learning and memory,spatial memory and other cognitive functions were weakened,and the escape latency was prolonged.The evasion latency of the positive drug group,the middle dose group and the high dose group were significantly shortened,indicating that the learning and memory function of the scopolamine model mice was significantly improved.The low dose group had the effect of improving learning and memory function,but did not achieve the effect of middle and high dose.The learning and spatial memory abilities of mice were observed through spatial exploration experiments.Mice in model group received scopolamine,and the cognitive function of mice learning and spatial memory decreased,and the number of crossing platform decreased.The number of cross platform in the low,middle and high dose group of JTW and positive drug group increased,which indicated that it had obvious effect on improving the learning and spatial memory ability of scopolamine model mice.The memory function of mice was observed in the space exploration test.The cognitive function of memory and spatial familiarity of mice in model group were weakened,and the residence time of space exploration experiment in platform quadrant decreased.The space exploration test in the low,middle and high dose group of the JTW and positive drug group increased in the platform quadrant,which indicated that it was effective for improving the memory function of scopolamine model mice.In the Y-maze test,the percentage of the spontaneous intercourse reaction in the scopolamine model group was significantly different from that in the control group,P < 0.05,indicating that the working memory function of the model group was lower than that of the control group.The percentage of spontaneous intercourse reaction in the positive drug group was compared with the model group.Although it had a trend of improvement,it did not have a statistical difference of P > 0.05.It showed that they had no obvious effect on improving the cognitive dysfunction of learning in mice.Compared with the model group in the low,middle and high dose group of JTW the percentage of spontaneous intercourse replacement was statistically different P < 0.05,indicating that the lower,middle and high dose groups could improve the memory cognitive function of mice.It can be seen that in improving the memory function of mice,the low,middle and high dosage of JTW can improve the memory function of mice.In the experiment of new object recognition,the identification index of new objects in the model group of scopolamine was lower than that in the control group,but it did not have statistical significance P > 0.05.There was no significant difference between the new thing identification index and the model group,but the trend of improvement was P > 0.05,indicating that the ability of improve the familiarity with the fresh things was improved,but it was not obvious.Compared with the model group,the low,middle and high dose group of JTW had no significant difference,but the trend of improvement was P > 0.05,indicating that the lower,middle and high dose groups improved the ability of the mice to be familiar with the fresh things.It can be seen that the low,middle and high dosage of JTW can improve the ability of mice to recognize and remember new things.In the condition of condition fear experiment,after the adaptation period,the freezing times of the scopolamine model mice was significantly different from that of the control group,P < 0.01,indicating that the memory function of the model mice was significantly lower than that of the control group.There was a significant difference in the relative ratio of the freezing times of the positive drug group and the model group,P < 0.01,indicating that amelioris the memory function of the fear in mice.Compared with the model group,the lower,middle and high dose group of JTW had a tendency to change the rigidity of the fear,but there was no statistical significance P > 0.05.It showed that the improvement of the low,middle and high dose groups improved the fear memory ability of mice but was not obvious.Therefore,the improvement of the memory function of mice,the low,middle and high dose of the JTW can improve the memory function of mice.Mice in model group were treated with scopolamine,and the level of Ach in mice decreased.The level of Ach in the positive drug group and the low,middle and high dose group of the treatment group showed significant improvement in the level of Ach in the scopolamine model mice.The activity of AchE in the cortex and hippocampus of mice increased.The activity of AchE in cortex and hippocampus of mice with positive drug group and JTW low,middle and high dose group decreased,indicating that JTW group was effective in inhibiting the AchE activity of scopolamine model mice.The activity of ChAT in mice was weakened.The ChAT activity of the positive drug group and the low dose,middle and high dose group of the treatment group increased,indicating that the ChAT activity of scopolamine model mice was significantly improved.The level of MDA in the hippocampus and cortex of mice increased.The levels of MDA in the cortex and hippocampus of the mice of the positive drug group,the JTW middle and high dose group were decreased,indicating that the group of JTW has a certain effect on the inhibition of the MDA level of the scopolamine model mice.The level of reactive oxygen species(ROS)in the cortex and hippocampus increased.The level of reactive oxygen species(ROS)in cortex and hippocampus of mice of JTW group and positive drug group decreased,indicating that JTW group could improve the effect of inhibiting the active oxygen(ROS)level of scopolamine model mice.The activity of SOD in the hippocampus and cortex of mice decreased.The positive drug group,the low,middle and high dose group of the treatment group increased the activity of SOD in the hippocampus and cortex of the mice.The expression of BDNF,NGF,SYN and Psd95 in the low,medium and high dose group of JTW improved.The neurons in the hippocampus CA1 area of the model group of the scopolamine model group showed a distinct atrophy,irregular arrangement and weak staining,suggesting that neurons were damaged and spread or died,and a large number of tigroid body were lost in these neurons.In the low dose group,the neurons in hippocampus CA1 area of mice were partially filled,arranged in regular pattern,and stained normally.In the middle and high dose group,the neurons in the hippocampus CA1 area of mice were more regular,and the staining was normal.The low,medium and high dose group of JTW inhibited the apoptotic protein,and the positive TUNEL staining slice were dark brown.In contrast,the positive cells in the cortex of scopolamine model mice increased and the background dark brown was deepened.No dark cells were found in the positive drug donepezil group.A small number of positive cells were found in both low and medium doses of JTW,but no positive cells were found in high dose group.Compared with the control group,the scopolamine model group’s related behavioral indexes,cholinergic index,oxidative stress index,neuronal improvement and apoptotic protein all decreased;the positive drug donepezil group mice cholinergic index,oxidation The stress index,neuronal improvement,and apoptotic protein levels were improved compared with the model group;the cholinergic index,oxidative stress index,and neuronal improvement were compared between the low,middle,and high dose groups of JTW and the scopolamine model group,and the apoptotic protein situation improved.Conclusion:JTW can improve learning,memory,spatial identification,new object recognition,conditioned fear and other behavioral abilities of mice.JTW can improve the related indexes of cholinergic system such as Ach,AChE and ChAT in mice.JTW can improve the indexes of oxidative stress system such as MDA,ROS,SOD and CAT in mice.JTW can improve the expression of BDNF,NGF,PSD95 and SYN proteins and apoptotic proteins in mice.The morphology,staining and arrangement of neurons in hippocampal CA1 area can be improved by JTW.JTW can inhibit and protect the apoptosis of mice nerve cells. |
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