| Objective: In this study,APP/PS1 transgenic mice were used as research objects to explore the effect and mechanism of exercise and RNSP in improving AD,and to compare the neuroprotective effect of RNSP and RNSP on AD,so as to provide theoretical guidance for exercise and RNSP in the prevention and treatment of AD.Methods: Twenty 16-week-old male wild mice were divided into WTE group and WTC group,with 10 mice in each group.Seventy 16-week-old male APP/PS1 transgenic mice were randomly divided into ADC group,ADE group,medium dose of original drug group,low dose of decoction group,medium dose of decoction group,high dose of decoction group and donepezil group.The exercise group and the medication group were given moderate intensity aerobic exercise intervention and gavage intervention for 10 weeks,respectively.After the intervention,open field test and Morris water maze test were performed to evaluate the cognitive and memory abilities of mice.At the end of the behavioral experiment,3 rats from each group were randomly selected for cardiac perfusion for brain harvesting,and the remaining rats were sacrificed under anesthesia,and the bilateral hippocampus was harvested.ELISA was used to detect the levels of Aβ,Bcl-2,Bax,ATP and AMP.HE staining was used to observe the state of hippocampal vertebral cells.The protein expression levels of AMPK,p-AMPK,HDAC5,p-HDAC5,HIF-1α,VEGF,H3K27 ac,and BDNF were detected by Western Blot.Immunofluorescence was used to detect the expression of HDAC5 in hippocampus.Results: 1.Open field experiment: Compared with WTC group,the total distance of ADC group decreased significantly(P<0.01);Compared with ADC group,ADE group,original medium dose group and reduced dose groups,the total distance was significantly increased(P<0.05).Compared with the WTC group,the movement time and The Times of entering the central area in ADC group were significantly decreased(P<0.01).Compared with ADC group,the central movement time and times of entering the central area were significantly increased in ADE group(P<0.01),and the central movement time and times of entering the central area were significantly increased in original medium dose group and reduced prescription dose groups(P<0.05).There were no significant differences between ADE group and drug groups in total distance,central area movement time and central area entry times(P>0.05).2.Morris Water Maze: During the positioning voyage,compared with the WTC group,the latency of the ADC group was significantly increased(P<0.01);compared with the ADC group,the latency of the ADE group,the medium dose of the original medicine group and the high dose of the reduced prescription group was significantly decreased(P<0.05);the latency of the medium dose of the reduced prescription group was significantly decreased(P<0.01).In space exploration,there was no significant difference in the total swimming distance of 60 s among all groups(P>0.05).Compared with the WTC group,The Times of crossing the platform and the percentage of the platform quadrant path in ADC group were significantly decreased(P<0.01).Compared with ADC group,ADE group,The Times of crossing the platform and the percentage of platform quadrant path in the low and high dose groups of the original medicine and reduced prescription were significantly increased(P<0.05),and The Times of crossing the platform and the percentage of platform quadrant path in the reduced prescription medium dose group were extremely significantly increased(P<0.01).There were no significant differences in latency,times of platform crossing and percentage of platform quadrant distance between ADE group and drug groups(P>0.05).3.ELISA detection: Compared with WTC group,the levels of Aβ and Bax in ADC group were significantly increased(P<0.01),while the levels of Bcl-2,AMP and ATP were significantly decreased(P<0.01).Compared with ADC group,Aβ levels in the medium-dose group of original medicine and low-dose group of reduced prescription were significantly decreased(P<0.05),Aβ levels in ADE group and medium-dose and high-dose groups of reduced prescription were significantly decreased(P<0.01),and Bax levels in ADE group,medium-dose group of original medicine and dose groups of reduced prescription were significantly decreased(P<0.05).The level of Bcl-2 was significantly increased(P<0.05),the level of AMP was significantly increased(P<0.01),the level of ATP in the medium-dose group of the original drug and the low-dose and high-dose groups of the reduced formula were significantly increased(P<0.05),the level of ATP in the ADE group and the medium-dose group of the reduced formula was significantly increased(P<0.01);There were no significant differences in the levels of Aβ,Bax,Bcl-2,AMP and ATP between ADE group and drug groups(P>0.05).4.HE staining: In the hippocampus of ADC group,a large number of pyramidal cells were necrotic,cell body disintegration and fragmentation were observed,the number of cell bodies was significantly reduced,the arrangement of pyramidal cells was disordered,nucleolar shrinkage and deep staining were observed,cell bodies became significantly smaller,and the internal structure of cells was unclear.Pyramidal cell necrosis was improved in ADE group,medium dose group and reduced dose group.5.Western Blot: there was no difference in the expression of AMPK protein in each group(P>0.05).Compared with WTC group,the protein expression levels of P-AMPK and P-HDAC5 in ADC group were significantly decreased(P<0.01),while the protein expression levels of HDAC5 were significantly increased(P<0.01).Compared with ADC group,ADE group,ADE group,medium dose group and reduced dose groups,PAMPK protein expression was significantly increased(P<0.05),ADE group,PHDAC5 protein expression was significantly increased(P<0.05),and P-HDAC5 protein expression was extremely significantly increased(P<0.01)in medium dose group and reduced dose groups.The expression of HDAC5 protein in ADE group and high-dose reduced prescription group was significantly decreased(P<0.05),and the expression of HDAC5 protein in medium-dose and low-dose reduced prescription groups was extremely significantly decreased(P<0.01).Compared with WTC group,HIF-1α protein expression in ADC group was not significantly different(P>0.05),but H3K27 ac protein expression was significantly decreased(P<0.05).Compared with ADC group,the expression of HIF-1α protein in ADE group and reduced prescription low-dose and high-dose groups was significantly increased(P<0.05),the expression of HIF-1α protein in original prescription medium-dose and reduced prescription mediumdose groups was extremely significantly increased(P<0.01),and the expression of H3K27 ac protein in ADE group was significantly increased(P<0.05).The protein expression of H3K27 ac in the medium dose group and the reduced dose groups was significantly increased(P<0.01).Compared with WTC group,the expressions of VEGFA and BDNF in ADC group were significantly decreased(P<0.05).Compared with ADC group,ADE group,original medium dose group and reduced dose groups,VEGFA and BDNF protein expressions were significantly increased(P<0.05).There were no significant differences in the protein expressions of AMPK,P-AMPK,HDAC5,p-HDAC5,HIF-1α,VEGF,H3K27 ac and BDNF between ADE group and drug groups(P>0.05).6.Immunofluorescence: Compared with WTC group,the percentage of HDAC5 positive area in ADC group was significantly increased(P<0.01),and the percentage of HDAC5 positive area in nucleus was significantly increased(P<0.05);Compared with ADC group,the proportion of HDAC5 positive area in ADE group,active dose group and reduced dose groups was significantly decreased(P<0.01),and the proportion of HDAC5 nuclear positive area in active dose group,reduced dose group and high dose group was significantly decreased(P<0.05).The proportion of HDAC5 nuclear positive area in ADE group and reduced prescription low-dose group was significantly decreased(P<0.01).Conclusion: 1.10-week exercise and reduced RNSP intervention can reduce the deposition of Aβ in the hippocampus of APP/PS1 mice,inhibit the apoptosis of hippocampal neurons,and improve the cognition and memory ability of mice.2.10-week exercise and reduced RNSP intervention can improve energy metabolism disorders in the hippocampus of APP/PS1 mice,and promote the nuclear export of HDAC5 by increasing AMPK activity,thus exerting neuroprotection.3.10-week exercise and reduced RNSP intervention could activate HIF-1α/VEGF and H3/BDNF pathways in the hippocampus of APP/PS1 mice through AMPK/HDAC5 pathway,thereby improving vascular function and promoting neuronal regeneration,and ultimately improving AD.4.There was no significant pharmacodynamic difference between original RNSP and reduced RNSP in improving AD pathology. |
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