The Effects Of Electricity On Learning And Memory In Rats With Chronic Alcoholism Were Studied Based On Metabolomics

objective Metabolomics was used to observe the effects of electroacupuncture at baihui and dazhui on hippocampal metabolism in rats with chronic alcoholism,and to find possible new mechanisms for electroacupuncture to treat learning and memory impairment caused by chronic alcoholism.Methods Adult SPF-grade SD rats were randomly divided into two groups: a blank control group and an alcohol model group.The blank control group was fed with an equal volume of normal saline at a rate of 10 ml / kg per rat.The alcohol model group used 56 The red star Erguotou was tested according to the standard of 10 ml / kg per rat.After 6 weeks of gastric administration,the learning and memory abilities of rats were measured by Morris water maze test.Rats with learning and memory impairment induced by chronic alcoholism were randomly divided into model group and electroacupuncture group,with 15 rats in each group,for a total of 45 rats.Six weeks after the model was established,electroacupuncture at Baihui and Dazhui was given.The treatment intensity was voltage 3V,current 1mA,frequency 3Hz,and continuous wave treatment for 30 min.After the experiment,the rats were decapitated and the brain was taken to separate the hippocampal tissue.The metabolites of the samples were extracted and chromatographically separated using an ultra high performance liquid chromatography system(UHPLC)using a HILIC column.Mass spectrometric analysis was performed to obtain a map of the differential metabolites,and then the XCMS program was used for peak alignment,retention time correction,and extraction of peak areas.The metabolite structure was identified by accurate mass matching(<25 ppm)and secondary spectrum matching.The laboratory's self-built database was searched.For the data extracted by XCMS,delete the ion peaks with missing values> 50% in the group,integrate the positive and negative ion peaks and apply the software SIMCA-P 14.1(Umetrics,Umea,Sweden)for pattern recognition.After the data is preprocessed by Paretoscaling,Multi-dimensional statistical analysis of data,including unsupervised principal component analysis(PCA)analysis,supervised partial least squares discriminant analysis(PLS-DA)and orthogonal partial least squares discriminant analysis(OPLSDA),and then significant difference metabolites were analyzed Identification and analysis,further bioinformatics analysis,screening of significantly different metabolites,and hierarchical clustering and KEGG metabolic pathway analysis were performed to summarize the analysis of the metabolic pathways involved;and the accuracy of the results by Western Blot technology for further verification.Results1.Morris water maze test results: After 6 weeks of modeling,the learning and memory ability of rats in the alcohol model group was significantly reduced,which was significantly different from that of the blank control group(P <0.05).Compared with the model group,the ability of learning and memory has improved,and there is a significant difference(P <0.05).2.Comparison of metabolic profiles of rat hippocampus tissues: There is a clear distinction between the model group and the electroacupuncture group,suggesting that the metabolite spectrum of rat hippocampus tissues changes drastically during the development of chronic memory-induced learning and memory impairment.Promote the metabolic profile of hippocampus of chronic alcoholism rats to normal levels.3.Electro-acupuncture for chronic alcoholism-related metabolic markers in rats: Based on multidimensional statistical analysis of VIP> 1 and univariate statistical analysis of metabolites with P value <0.05,differential metabolites were screened including creatine,glutamic acid,and glutamine amber Acid,histidine,methionine,?-ketoglutarate,glyceryl phosphate choline,fumaric acid,taurine,etc.These metabolic markers have dynamic changes in electroacupuncture intervention in rats with chronic alcoholism-induced learning and memory impairment,Suggesting that in the occurrence and development of energy metabolism,antioxidant immunity,cell activity and neurotransmitter disorders.Electroacupuncture has different degrees of regulation on metabolic markers.4.Effects of electricity on metabolic pathways in hippocampus tissue of chronic alcoholism model rats: mainly involves 10 pathways,mainly involved in energy metabolism,amino acid and amide metabolism,mainly including the metabolism of glycine,serine and threonine,alanine,Aspartic acid and glutamic acid metabolism,as well as neurotransmitter signaling,protein digestion and absorption,glycerophospholipid metabolism,amino acid biosynthesis,aminoacyl-tRNA biosynthesis,abc transporter.The differential expression of selected proteins was observed by western blot,and the change trend was consistent with metabolomics results.Conclusion Electroacupuncture stimulation of baihui point and daji point in rats with chronic alcoholism may regulate and protect the learning and memory ability by affecting the glutamate metabolism pathway in the hippocampus.