A Comparative Study On 5hmC Targeting Regulation Of Neurons In AD Mice By Several Natural Compounds

Alzheimer’s disease(AD)is a progressive neurodegeneration disease that usually results in selective neuron loss and degeneration,amyloid plaque(SP)deposition,neurofibrillary tangles(NFT)in the hippocampus,cortex,and other tissues,even leads to nerve death.Due to the complex pathogenesis,there are no effective prevention and treatment measures at present.AD has become a major threat to the health of old people.In recent years,studies have found 5-methylcytosine(5mC)can be oxidized to5-hydroxymethylcytosine(5hmC)by the TET family proteins.As a key intermediate for DNA demethylation,5hmC plays an important role in maintaining the normal physiological function of the body.A series of studies have confirmed the disorder of DNA methylation(5mC/5hmC)is closely related to the occurrence and development of the diseases,and the deregulation of DNA methylation do exists in neurodegenerative diseases,such as AD.This study conducts research on cell-level,selecting five natural compounds(cordycepin,cordycepic acid,geniposide,crocin,and ergosterol)that have been proven to improve memory,neuroprotection,and enhance immunity as research materials,and chose the resveratrol as the positive control.It intends to discover natural compounds which can adjust and control 5hmC levels and improve AD neuronal status.The results of Dot Blot showed that cordycepin,cordycepic acid,resveratrol and crocin can increase the expression level of 5hmC significantly.Combined with qPCR results,we found that cordycepin can increase the expression of Tets mRNA compared with the above-mentioned natural compounds.Also,it can reduce the transcription level of ApoE significantly,suggested that cordycepin may reduce the formation of NFTs and the accumulation of Aβ in the brain by reducing the expression of ApoE,thereby affect the progression of AD.Meanwhile,the immunofluorescence staining results showed after treatment of cordycepin and corcycepic acid,the percentage of differentiation of SHSY-5Y cells increased significantly.Cordycepin can increased the length of axons and the number of dendritic branches in mouse primary neurons significantly at thesame time.The screening results showed that cordycepin had a positive influence on the level of 5hmC or on the morphology of neurons,and most of the effects were better than the positive control(resveratrol),indicating that cordycepin may delay the progression of neurodegenerative diseases such as AD,but the specific mechanism of action remains further study.Our research provided a basis for further discussion on the influence of cordycepin on AD and a new idea for the pathological study of related diseases.