| Objective: Alzheimer's disease?AD?is a progressive neurodegenerative disease,is the most common type of dementia.The deposition of amyloid beta?A??is still considered as a hallmark pathological feature of the disease.AE-624 is one of the active ingredients of phenylethanoid glycosides in Cistanches Herba.Previous studies have confirmed that AE-624 can improve the learning and memory impairment in a variety of AD mouse model.This study will examine the effects of AE-624 on the aggregation of A?in vitro at the molecular level;To investigate the protective effect of AE-624 on A?induced injury of PC12 cells at the cellular level;APP/PS1 double transgenic mice were selected as AD mice model;To investigate the effect of AE-624 on the cognitive behavior of APP/PS1 double transgenic mice,the function mechanism of the prevention and treatment of AD.Method:?1?40?mol/L CQ as positive drug,using thioflavin T?Th-T?fluorescence detected the effect of?40,80,160?mol/L?AE-624 on the monomer A?1-42 and the aggregated A?1-42,?2.5,5,10,20,40,80,160,320,640,1280,2560?mol/L?AE-624 present a Does effect relationship;In order to rule out the possible false positive results in the experiment,300600nm full wavelength absorption spectrum scanning for AE and AE+Th-T,and competitive binding assay was applied to?160,320,640?mol/L?AE-624 and?1.25,2.5,5,10,20,40,80,160,320?mol/L?Th-T.PC12 cells were treated with 40?mol/L A?1-42,the drug group were added with?1,10,100 ?mol/L?AE-624,incubated for 24 h,the survival rate was measured by MTT assay.?2?9-month old APP/PS1 mice by nesting experiment and weight were divided into model group,donepezil group?3mg/kg?,AE-624?20,40,80mg/kg?group,the normal control group was9 months old C57BL/6J mice.Nesting experiment once a week,After intragastric administration of 134 d,through aging score,nesting score,autonomic activities,new object recognition experiments,O maze and Morris water maze test,shuttle box test,toobserve the effect of AE-624 on aging degree,nest building behavior,autonomic activity,anxiety,learning and memory ability of APP/PS1 mice;Through A? Immunofluorescence staining in the cortex and hippocampus of mice and detection of A?1-40 in hippocampus of mice by Elisa,to observe the effects of AE-624 on A? deposition in APP/PS1 mice;Observe the effect of AE-624 on CA1 and cortical region neurons loss of mice by Nissl staining;?3?Observe the effect of AE-624 on HPA,HPG,HPT and HPGH axis hormone secretion and BDNF expression level and peripheral blood cytokines secretion influence of APP/PS1 mice by Luminex analyzer;Observe the effect of AE-624 on oxidative factors by measuring the SOD inhibition rate and MDA,GSH-Px activity in APP/PS1 mice blood;Using Pearson correlation analysis method,to investigate the correlation of cytokines,hormones,Oxidation factor in NIM network respectively with APP/PS1 mice aging degree,Non-cognitive behavior,cognitive function,selection of seed gene to establish PPI network.Result:?1?Thioflavin T binding experiments show that add?160?mol/L?AE-624 co incubated after 24 h,the monomer and the aggregated A?1-42 fluorescence intensity significantly decreased,After excluding false positive results,Effects of AE-624 on the dose effect relationship of A?1-42 aggregation,AE-624 inhibit A?1-42 aggregation,the IC50 were 254.3?mol/L,AE-624 depolymerization the aggregated A?1-42,the IC50 was282?mol/L.For A?1-42 induced PC12 cell injury,100?mol/L AE-624 significantly increased cell viability?P<0.01?.?2?AE-624?20,80mg/kg?can effectively control the weight of APP/PS1 mice?P<0.05?;improved APP/PS1 mice nest score trend,no significant difference;Aging scores showed that,compared with the model group,AE-624?20,40,80mg/kg?groups can slow the aging process of APP/PS1 mice?P<0.001,P<0.01,P<0.05?;Compared with the model group,AE-624?80mg/kg?group can weaken the manic mood of mice?P<0.05?;O maze showed that AE-624?20mg/kg?group can alleviate the anxiety of mice?P<0.05?;In the new object recognition experiment,AE-624?40mg/kg?group had a significant effect on the short-term memory of mice?P<0.05?;Morris water maze showed that AE-624 had no significant effect on spatial memory in APP/PS1 mice;AE-624?40mg/kg?group has the ability to improve the learning disabilities of the model mice in the shuttle box test;Compared with the normal group,the content of A?1-40 in the hippocampus of APP/PS1 mice were significantly increased,the number of Nissl bodies in the CA1 and cortical areas of hippocampus was significantly reduced and the cell arrangement was sparse,AE-624?40mg/kg?significantly decreased the content of A?1-40?P<0.01?in the hippocampus of APP/PS1 mice,decreased the deposition of A beta,increased the number of Nissl bodies in CA1 andcortex.?3?AE-624?80mg/kg?can significantly decrease the ACTH and CORT contents in HPA axis of APP/PS1 mice?P<0.05?;AE-624?20mg/kg?inhibited mouse HPG axis on the secretion of FSH?P<0.05?,AE-624?80mg/kg?can significantly increase the TSH content of HPT axis in APP/PS1 mice?P<0.01?,The GH secretion of HPGH axis was improved,and the deficiency of BDNF in the mouse pituitary was effectively improved?P<0.05?;AE-624 can significantly reduce the chemokines in peripheral blood of mice?eotaxin,RANTES?content,and improve the content of anti-inflammatory factor G-CSF in blood;AE-624 can increase the inhibition rate of SOD and the activity of GSH-Px,decreasing the content of MDA?P<0.001?;By Pearson correlation analysis,Get the hormone,cytokines,oxidative factors,Which was significantly associated with the aging,non-cognitive behavior and cognitive behavior of APP/PS1 mice,Suggesting that these molecules may be AE-624 potential target for the prevention and treatment of AD.These were used as seed genes to construct the PPI network,The 56 nodes involved in the network are imported into the DAVID server,Through P Value,we screened out three significant enrichment related channels of AE.Conclusion:?1?AE-624 can effectively inhibit the aggregation of the monomer A?1-42,also has the obvious depolymerization on the aggregated A?1-42 in vitro.AE-624 has significant protective effect on PC12 cells injured by A?1-42?2?Long-term administration of AE-624 can effectively control the body weight of APP/PS1 mice,delay aging,improve the anxiety,improve memory function of mice.?3?AE-624 can improve APP/PS1 mice HPA,HPG,HPT,HPGH axis disorder,improve mouse brain derived neurotrophic factor BDNF deficiency,improve the anti-inflammatory factor in blood and chemokine levels,by improving the antioxidant capacity of ways to prevention and cure senile dementia;AE-624 mainly act on CORT,BDNF,MDA,GSH-Px to delay the aging of APP/PS1 mice,act on SOD and MDA,BDNF,ACTH and G-CSF to improve non cognitive,act on MDA,CORT,LH,BDNF,GH and TSH to improve cognitive behavior in mice,and find out the possible pathway of AE-624. |
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