Exploration On The Influence Of Acetylcholinesterase Inhibitors On Regulating Lipid Metabolism For Alzheimer's Disease Based On Traditional Chinese Medicine Theory "Xin Zhu Shen Zhi"

Alzheimer's disease(AD) is a complex neurodegenerative disorder characterized by a multi-factorial etiology not completely understood and lacking early biochemical diagnosis.Donepezil(DNP) as the first-line acetylcholinesterase inhibitors in treatment of mild-to-moderate AD Patients have a different response to DNP and the therapeutic response to DNP could not be evaluated.DNP was found to have other non-cholinergic mechanisms besides potent acetylcholinesterase inhibitory effect,while the mechanism is undefined.Our goal of this study is to elucidate the potential role of AD and donepezil therapy on lipid metabolic parameters,simultaneously,discover lipid candidate diagnostic and therapeutic biomarkers.In the opinion of western medicine,the main pathological changes occurred in brain and the studies focus on brain as the target.This study was based on the traditional Chinese medicine's theoretical system as "Xin Zhu Shen Zhi”(close relationship between heart and brain,heart governs mind).The physical base of the traditional Chinese medicine was blood including the small molecular energy substance and regulators of energy metabolism.Traditional Chinese medicine treatment based on this theory had a good effective on AD treatment.We design our study according to this theory and focus on the relationship of heart and brain.Since ketone body and adiponectin acts the same role in heart and brain during the progress of the disease and based on the traditional Chinese medicine's theoretical system as "heart governs mind",we focus on adipokine and small molecular lipids differences between AD and DNP treatment.In the present study,we divided AD patients newly treated with DNP into responder and non-responder groups.We applied non-target and target metabolomics approach to characterize metabolic alterations of different groups including AD patients,controls,DNP responder and non-responder.Metabolomics analyses were carried on GC-MS.The metabolite classes that were found to be significantly altered between control,AD,DNP-R and DNP-N include plasma lipid metabolites and amino acids,branch-chain amino acids.Compared to the control group,?-hydroxybutyrate was decreased in AD group.DNP treatment ameliorated the trends of the decrease of ?-hydroxybutyrate.In order to identify whether DNP treatment ameliorates the lipid metabolic pattern in AD,the target metabolomic of ?-hydroxybutyrate was evaluated by LC-MS/MS.This targeted analysis revealed significantly lower plasma levels of ?-hydroxybutyrate in newly diagnosed AD group compared to the control group.DNP significantly elevated plasma ?-hydroxybutyrate levels when compared to AD group.The result indicated that DNP can affect the ?-oxidation-related lipid metabolism.Adiponectin as the regulator of lipid metabolism were assayed by Elisa Kits.Genotyping of adiponectin was performed using PCR-LDR ligase detection reaction.Plasma adiponectin levels were increased in AD group compared to the control group.Metabolomic data has also demonstrated profound impairments in pathways related to fatty acid oxidation,gluconeogenesis and lipolysis,and we identified the measurable metabolites in these selected pathways indicate decreasing fatty acid oxidation,increasing gluconeogenesis and stimulation of lipolysis was occurred in AD group.The level of adiponectin was elevated but the physiological effects were suppressed.The concept of adiponectin resistance maybe exists for the compensatory elevated adiponectin levels in AD patients.Plasma adiponectin levels were significantly decreased in DNP-R group compared to the DNP-NR group.We did not find a significant difference in adiponectin genotype between responders and non-responders of DNP.Adiponectin polymorphisms were not related to plasma adiponectin levels in our study.The effect of the DNP on the ketogenesis and adiponectin secretion was investigated in mice.Fasting animal and multiple-dose oral administration experimentation was carried to evaluate the effect of DNP on ketogenesis and adiponectin secretion.We found DNP was capable to readdress ketone body production.?-hydroxybutyrate level in serum and liver from DNP treated mice exhibited a dramatic increase compared to control.Also,the key enzymes involved in ketone body production,such as HMGCS1,HMGCL and BDH are clearly upregulated in DNP treated group,relative to untreated groupBased on the traditional Chinese medicine's theoretical system as "heart governs mind",we investigated the effect of DNP on ketone body utilization in H9C2 cardiomyocyte.We found glucose induced changes in the expression of 3-oxoacid CoA transferase and DNP could upregulate OXCT.We have demonstrated that high glucose caused OXCT expression downregulated was reversed by DNP and DNP could increase the utilization of ketone body in cardiomyocyte.In summary,in the present study,we have observed striking differences in the serum levels of adiponectin and ?-hydroxybutyrate in AD subjects compared to control,which are likely to have important implications in the disease pathogenesis and could be potential biomarkers of disease diagnosis.Our studies highlight findings underline the important of DNP in the regulation of ketogenesis and ketolysis.These findings are important for clinical practice.The study was guided by the Chinese medicine theory,and the results also confirmed the scientific nature of traditional Chinese medicine theory.