The Therapeutic Effect Of Zhenbao Pills On Neuropathy In Zebrafish With Imitated Alzheimer’s Disease

Zhenbaopill（ZBP）is a traditional clinical Mongolia medicine with 241 active ingredients as indicated by analysis and is mainly used in clinical practice for the treatment of neurological disorders.Alzheimer’s disease（AD）is a common neurodegenerative disease.The purpose of this study was to investigate the mechanism of action by evaluating the therapeutic effect of ZBP on an Al Cl₃trichloride-induced imitate zebrafish AD model.In this study,we first established a imitate AD zebrafish model induced by Al Cl₃exposure and tested the therapeutic effect of ZBP on the AD zebrafish model after treatment by the effect of different concentrations of ZBP.The main test indexes included behavioral tests,such as swimming distance and swimming speed of zebrafish,and reactivity to determine the recovery of behavioral impairment on the AD zebrafish model.Histology was determined by HE staining of the brain and TUNEL（terminal deoxynucleotidyl transferase d UTP nick-end labeling）staining to determine pathological changes in the brain of the AD zebrafish model.The pathways of action of ZBP were then determined by transcriptomic analysis and the results were validated by fluorescent quantitative PCR.Finally,the pathway of action of ZBP was revalidated by knockdown and overexpression techniques.The experimental results revealed that Al Cl₃treatment caused the behavioral impairment,induced apoptosis in the brain,produced apoptotic vesicles,increased acetylcholinesterase（ACh E）activity,and decreased acetylcholine（ACh）content in zebrafish.However,treatment with ZBP at 16μg/m L was effective in alleviating behavioral disturbances in zebrafish larvae,an effect superior to that of the cholinesterase inhibitor Donepezil Hydrochloride（DPZ）.Transcriptomic data analysis showed that ZBP mainly acted on apoptosis,p53 signaling pathway,and enrichment of ACh E-related pathway.Furthermore,overexpression of ACh E m RNA produced a similar AD-like disorder,whereas ACh E knockdown（ACh E-MO）reduced ACh E activity and alleviated AD-like symptoms.These results suggest that ZBP is a potential cholinesterase inhibitor and portends great developmental value for the use of ZBP in the treatment of AD.