Pharmacodynamics And Mechanical Study Of Alpiniae Oxyphyllae Fructus In The Treatment Of Alzheimer's Disease Based On Metabolomics And Network Pharmacology

Objectives:To explore the endogenous differential metabolites and the metabolites reversed by the total extract of AOF related to the occurrence and development of Alzheimer's disease rats,and to construct the metabolic regulation network of the total extract of AOF,and to explore the "multiple components-multiple targets-multiple pathways"mechanism of AOF total extract in the treatment of Alzheimer's disease from two aspects of metabolism and protein targets.Methods:1.Based on ultra high performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrap high resolution accurate mass spectrometry(UHPLC-Q-Orbitrap HRMS)technology,the chemical constituents of AOF total extract were studied,and the compositions of complex chemical components in AOF were identified.2.Through the establishment of AD rat model,the behavioral changes and the relative content of A? protein in brain were observed by water maze test and immunohistochemistry,and then the protective effects of AOF total extract on AD rats were evaluated.3.Through the establishment of non-target metabolomics,the endogenous differential metabolites of AD and the negative regulatory metabolites of AOF were found.Based on these metabolites,the metabolic regulatory network of AOF total extract was constructed,and the action mechanism of AOF extract on AD rats was discussed from the metabolic level.4.Based on the method of network pharmacology,the potential action targets of AOF were predicted by using a large number of databases with potential small molecular components as probes.By comparing the predicted target with the related target of AD metabolites that can be recalled by AOF extract,we found the core target of AOF total extract in regulating Alzheimer's disease,and further elucidated the mechanism of "multiply components-multiply targets-multiply pathway" of AOF total extract on AD.By comparing the predicted target with the related target of AD metabolites that can be recalled by AOF extract,we found the core target of AOF total extract in regulating Alzheimer's disease,and further elucidated the mechanism of "multiple components-multiple targets-multiple athways" of AOF total extract on Alzheimer's disease.Results:1.A method was successfully established for the determination of chemical components of AOF total extract based on UHPLC-Q-Orbitrap HRMS technology.A total of 48 chemical constituents were identified,including sesquiterpenes,flavonoids,diphenylheptanes and other components.2.The rat models of AD were successfully established by injecting A?1-42 protein into the hippocampus.The AOF can improve the behavior,memory and spatial cognition of AD rats.The results of immunohistochemistry showed that there were obvious A?1-42 yellowish brown positive plaques in the cerebral cortex and hippocampus in the model group compared with the sham operation group,and the area of A?1-42 yellowish brown positive plaques in the AOF group was significantly reduced compared with in the model group.Through the statistical analysis of the relative content of A?1-42 protein,it was found that the degree of reduction was statistically significant.3.A non-targeted metabolomics method was established for the treatment of AD with total extract of AOF by UHPLC-Q-OrbitrapHRMS technology.29 differential metabolites in plasma and 11 differential metabolites in brain tissue related to AD were identified.In addition,23 plasma and 9 brain tissue metabolites were identified,which were regulated by AOF.They mainly played a therapeutic role through amino acid metabolism,lipid metabolism and energy metabolism.4.Based on the network pharmacology method,it was found that AOF could regulate the metabolic disorder of AD by acting on 27 protein targets,such as FAAH,PPARG,ACHE BCHE,ADH1C and et al.Based on these targets,17 potential therapeutic substances were found,mainly including nootkatone,yakuchinone a,yakuchinone b,izalpinin,chrysin,etc.And the "multiple components-multiple targets-multiple athways" mechanism of the AOF in the treatment of AD was revealed.Conclusions:The multiple components of AOF could coordinate to regulate the metabolic disorder of AD,mainly involving amino acid metabolism,lipid metabolism,energy metabolism,etc.Through the integration of metabolomics and network pharmacology,the "multiple components-multiple targets-multiple pathways"mechanism of AOF in the treatment of AD was preliminarily explained from the metabolic level and protein target level,which provided a molecular basis for the treatment of AD,and provided new insights and theoretical support for other related research.