Preliminary Study On The Effect And Mechanism Of Tu-Xian Mixture On Diabetic Cognitive Impairment In Rats

[Objective]Traditional Chinese medicine(TCM)has long been used in the treatment of diabetic cognitive impairment(DCI),and Tu-Xian Mixture(TXM)is one of the prescriptions.In this study,the research methods of network pharmacology,gut microbiota and metabolomics were used to explore the effect and mechanism of TXM in treating DCI rats.[Methods]1.Search the database and consult the relevant literature to obtain the components of the five Chinese herbs in TXM and screen them according to oral bioavailability(OB)?30%and drug-likeness(DL)? 0.18.Query the targets of the selected components and the disease target of DCI from the database,and conduct KEGG pathway analysis together.2.40 SPF SD rats were divided into normal group(Con group,n=10)and model group(n=30)according to the random number table method.The rats in the model group were fed with high-sugar and high-fat diet for 6 weeks and then injected with STZ solution at 35mg/kg intraperitoneally.After being modeled,they were divided into diabetes model group(DM group,n=10),TXM group(Txm group,n=10),and donepezil group(Als group,n=10)according to the random number table method.The Txm group was given TXM 5g/(kg·d)by gavage,the Als group was given donepezil 0.5g/(kg·d)by gavage,and the Con group and DM group were given the same amount of distilled water once a day for 10 weeks.Measure body weight every week,and check blood glucose every two weeks.Morris water maze was used to detect the cognitive function of rats,Nissl staining was used to observe the morphology and number of hippocampal neurons,and Western Blot was used to detect the expression of BDNF and GABA A receptor.3.Collect rat feces,extract fecal genes for 16SV3-4 region sequencing.Analyze the difference in the structure of gut microbiota between normal rats and DCI rats,and the influence of TXM on the gut microbiota of DCI rats.4.Based on UPLC-MS/MS broad-target metabolomics,PCA and OPLS-DA were used to identify the metabolites in the hippocampus tissue of different groups of rats,analyze the difference metabolites and construct the KEGG pathway analysis.[Results]1.A total of 51 active ingredients and 619 potential targets were obtained,among which there were 33 key targets for diabetes and 18 key targets for cognitive impairment.Steroid hormone biosynthesis and y-aminobutyric acid signaling pathway may be the important metabolic pathways for TXM to perform therapeutic effects.2.TXM intervention reduced the latency period(p<0.05),and increased the number to cross the platform area(p<0.05)and the time stay in the target quadrant(p<0.05)in the Morris water maze.TXM improve the damage of high glucose to hippocampal neurons,reduce the apoptosis of them,up-regulate the expression of BDNF(p<0.05)and GABA A receptor al(p<0.05).3.After the intervention of TXM,6 kinds of intestinal microorganisms were significantly enriched in the feces of DCI rats,among which 4 kinds of microorganisms have been reported to be related to cognitive function.4.After the intervention of TXM,there were 24 differential metabolites in the hippocampus of DCI rats,13 up-regulated and 11 down-regulated.Among them,13 metabolites have been recalled,and 2 have been reported to be related to cognitive dysfunction(up-regulation of 3-hydroxybutyrate and down-regulation of malonic acid).KEGG pathway analysis showed that TXM may improve the cognitive function of DCI rats by regulating glucose and lipid metabolism,amino acid metabolism,ketone body synthesis and ferroptosis.[Conclusion]TXM can improve the cognitive ability of DCI rats,possibly by protecting hippocampal neurons,up-regulating the expression of BDNF and GABA A receptor ?1,correcting the gut microbiota disorder of DCI rats,and up-regulating 3-Hydroxybutyrate,down-regulating malonic acid,regulating glucose and lipid metabolism,amino acid metabolism,ketone body synthesis and degradation,and ferroptosis.[Innovation]1.The purpose of this study is to treat DCI,drawing on past clinical experience and medical research results to form a basic prescription TXM.2.Using the research methods of network pharmacology,gut microbiota and metabolomics,the preliminary exploration of the effect and mechanism of TXM in the treatment of DCI rats has been completed,and it also provided an experimental basis for the clinical application of TXM in the treatment of DCI.