Study On The Mechanism Of Action Of Wheat Embryo Globulin Fortified Complex In AD Model Rats

Alzheimer’s disease（AD）is a neurodegenerative disease characterized by decreased executive ability,visual impairment,abnormal mental action,and cognitive decline.With the aging of the world population,the prevalence of AD will continue to increase.At present,there is no effective method to treat or significantly improve AD,which brings a heavy burden to individuals and families.Therefore,focusing on the pathogenesis of AD,it is of great significance to study new therapeutic targets and intervention methods.The animal model of AD induced by D-galactose（D-gal）and aluminum chloride（AlCl₃）can effectively show all aspects of AD,such as pathological characteristics:β-Amyloid（Aβ）,Neurofibrillary tangles（NFTs）,and neuronal cell death,which are simple,effective and cheap,have been widely used by researchers.It is found that the combination of multiple nutrients can play its role better than a single component.The nutritional method of using the complex of multiple nutrients to prevent,delay or prevent the progress of ad is a promising strategy.Wheat embryo globulin（WEG）can improve the level of oxidative stress,inflammation,immune system damage,intestinal microbial disorder,and memory of cognitive impairment mouse model.Wheat embryo globulin fortified complex（WEGFC）is a functional food to strengthen WEG.The complex takes WEG as the main component,taurine,phosphatidylserineγ-Aminobutyric acid,vitamin B₁₂,folic acid,vitamin C,and vitamin E are a kind of functional food as auxiliary components.These auxiliary components contribute to the improvement of memory and is recognized as the main raw material of functional food that can improve memory.WEGFC is expected to become a functional food with potential intervention in AD.In this study,D-gal combined with AlCl₃ was used to induce AD rat model,and WEGFC was used to intervene the model rats.Through the research methods of behavior,pathomorphology,and molecular biology,this paper explores the protective effect and possible mechanism of WEGFC on cognitive impairment in AD rats,to provide theoretical support for the application of WEGFC as a functional food to alleviate cognitive impairment in AD.The main research contents and results of this paper are as follows:（1）Effect of wheat embryo globulin reinforcement complex on learning and memory in AD ratsThe AD rat model was established by D-gal combined with AlCl₃.Donepezil and WEGFC were divided into groups and intervened.The food intake and water intake were weighed and recorded,and the organ coefficients of brain,hippocampus,liver and kidney were calculated.The results showed that D-gal combined with AlCl₃ administration could lead to pathological state,weight loss,significant increase in food intake and drinking water per100 grams of body weight,and decrease in brain coefficient（p<0.05）.After supplementing of WEGFC,this situation was reversed and the body state of rats could be significantly improved,indicating that WEGFC can repair the physiological state of rats,It can also delay organ aging and enhance immune function.The learning and memory ability of rats was analyzed by water maze experiment,new object recognition experiment,and open field experiment.The results showed that WEGFC could improve the number of times of rats crossing the platform,residence time in the target quadrant,discrimination index of new objects,number of crossing grids and number of upright times（p<0.05）,and reduce the escape latency and number of fecal particles（p<0.05）,It shows that WEGFC can improve the decline of learning and memory ability,exploration ability,and tension and anxiety in rats caused by D-gal/AlCl₃.In this part,WEGFC（300 mg/kg）performed better in physiological indexes and behavioral experiments than donepezil group and WEG group,and the positive intervention effect was more significant,indicating that the function of WEGFC has been strengthened.（2）Effects of wheat embryo globulin fortified complex on biochemical indexes of AD ratsHE and Nissl staining were used to observe the hippocampal neurons and Nissl body of rats.The results showed that WEGFC reduced the damage of hippocampal neurons and increased the number of neurons（p<0.05）.It was found that WEGFC significantly increased SOD activity and GSH content in serum,cortex,and hippocampus,significantly reduced MDA content（p<0.05）,significantly increased Ch AT activity and ACh content in hippocampus,and significantly reduced Ach E,IL-1β,IL-6,and TNF-α（p<0.05）.Analysis of inflammatory signaling pathway:WEGFC significantly reduced TLR4/MyD88/NF-κB m RNA level and protein expression（p<0.05）.In this part,compared with non fortified WEG,the number of neurons,oxidation index,choline index,inflammatory index,and metabolic pathway of rats in WEGFC（300 mg/kg）or WEGFC（600 mg/kg）group are closer to those in the control group,which further shows that the function of wegfc intervention in AD model rats has been strengthened.（3）Effects of wheat embryo globulin fortified complex on intestinal microorganisms in AD rats16S rDNA high-throughput sequencing was used to analyze the changes of intestinal flora of rats in each group and the effect of WEGFC on intestinal microecology of AD model rats:WEGFC has little effect on the diversity and richness of intestinal flora of rats,which can increase the number of OTU shared with the control group and increase the similarity with the control group.At the door level,At the phylum level,WEGFC decreased the ratio of Firmicutes/Bacteroidetes associated with inflammation（p>0.05）.At the genus level,WEGFC significantly increased the abundance of beneficial flora Lactobacillus,a beneficial flora that can improve memory,and decreased the abundance of harmful flora Erysipelatoclostridium,Erysipelotrichaceae,Corynebacterium,and Frisingicoccus associated with inflammation（p<0.05）.Oxidative stress,neuroinflammation,cholinergic disorder,and intestinal microbial imbalance are the main theories of the pathogenesis of AD,NF-κB pathway is closely related to these theories,which can promote the pathogenesis of AD and form Aβdeposition,NFTs,and neuronal cell death eventually lead to cognitive impairment.Through the analysis of behavioral experiment,pathological experiment,oxidation index,choline index,and inflammatory index,the effect of wegfc group on AD model rats is more significant than that of WEG group,indicating that WEGFC has strengthened the function of improving cognitive impairment model rats.Anyway,we used D-gal/AlCl₃ to establish a rat model of learning impairment,and investigated the neuroprotective effect of wegfc from the physical characteristics,pathological characteristics and pathogenesis of AD.The results show that WEGFC improves the weight loss and learning and memory ability of AD model rats,alleviates the pathological damage of hippocampal tissue of AD model rats,improves the ability of antioxidant,neurotransmitter regulation and anti-inflammatory,increases the proportion of beneficial bacteria and reduces the proportion of harmful bacteria,and then plays a multiple protective role in improving cognitive impairment,which may be related to the activation of TLR4/MyD88/NF-κB signal pathway,which provides an experimental basis for wegfc as a functional food.