Preliminary Study On The Relationship Between The Regulation Of Gut Microbiota And The Improvement Of Insulin Resistance In T2DM By Chickpeas

ObjectiveType 2 diabetes mellitus(T2DM)is a wide spread metabolic disorder disease characterized by hyperglycemia.while the pathogenesis of T2DM is still unclear.Recently,although accumulating evidence indicates that the strong association between gut microbiota and T2DM,it’s still unclear between changes in gut microbiota caused by T2DM and changes in host’s endogenous metabolites.Chickpea(Cicer arietinum L.)has been long consumed as a kind of nutritional and health food in many countries.In addition to culinary usage,chickpea has been accepted as a natural Uighur traditional medicine in Xingjiang for a long time.Officially,chickpea is recorded as a treatment for T2DM due to its effective in improving T2DM.However,the mechanism of how chickpea improve T2DM is still unknown.Therefore,in this paper,we investigated that chickpea could alleviate T2DM via interactions with the gut microbiota and rebalance the disorder metabolism,which will provides a preliminary study of the mechanism of chickpea treatment of T2DM.Method1.Optimize the extraction methods of chickpeas and preparation of chickpea samples.Through six factors,including temperature,time and solvent,we chose 4 important factors,including ethanol concentration,temperature,time and the material-liquid ratio,and each factor was test 3 levels.Finally,we chose the best extraction method from total 29 BoxBehnken tests.2.To investigate the effect of chickpea on alleviating insulin resistance model rats with T2DM,we used the combination with high-fat diet and STZ intraperitoneal injection to established a T2DM rat model.Then,we gave the model rats with chickpea treatment,while using metformin as a positive control.During the experiment,the body weight and the changes of fast blood glucose were measured.In addition,the urine,fecal,blood and other tissue samples were collected.Subsequently,a series of indicators including blood sugar,blood lipids,immune factors,antioxidants and other indicators were measured.At the same time,the organ index was measured for the collected tissue samples and pathological sections were made.3.NMR was used to explore the metabonomic characteristics after chickpea in T2DM model rats.The metabolic pathway changes after chickpea changes were analyzed via urine and fecal samples.Biomarkers were selected by multivariate statistical analysis.Finally,explained the meaning of biomarkers using KEGG and Metabo Analyst.4.Based on 16S high-throughput sequencing technology to explore the intestinal microecological characteristics of chickpea in improving insulin resistance model of T2DM.After chickpea treatment,the sequencing gene of V4 region was analyzed the changes in gut microbiota after chickpea treatment.Alpha and beta diversity and LEfSe analysis were used to investigate structural changes in the host gut flora following chickpea intervention.The PICRUst function is then used to predict and analyze the affected path.Finally,the results of the PICRUst function prediction analysis are verified by the shotgun method.Result:1.Based on the content of isoflavones in chickpeas,the results showed that the optimum technological method was follows:ethanol concentration 70%,liquid-solid ratio 30:1,extracting temperature 80℃ and extraction time 1 h.In this case,the extracting of isoflavone reaches 1.702 mg/g under these conditions.2.From the analysis results of various physiological and biochemical indicators,it was found that chickpeas were used in T2DM model rats.Effectively reduce blood sugar,blood lipids,relieve inflammation,improve organ function and other aspects have been improved.3.Metabolomics analysis showed that amino acids(ornithine,arginine,aspartame)in T2DM model rats.Acid,lysine,methionine,glutamic acid),nucleotides and their metabolites(uracil,adenine,hypoxanthine),tricarboxylic acid cycle intermediates(citric acid,ketoglutaric acid,succinic acid)The contents of co-metabolites(short-chain fatty acids,taurine and hypotaurine)in the intestinal flora were significantly down-regulated,while the contents of glucose,phenylalanine,phenylacetic acid and hippuric acid were up-regulated.It is suggested that the amino acid metabolism,glucose metabolism and energy metabolism of T2DM model rats have different degrees of disorder,but they are corrected to some extent after the intervention of chickpea.4.Analysis of key populations of chickpea effect in fecal samples by 16s rRNA sequencing and shotgun sequencing treatment significantly improved hyperglycemia and inflammation in T2DM rats,similar to metformin.In addition,chickpeas can improve intestinal dysbiosis in T2DM rats,such as increasing the abundance of SCFAs and lactic acidproducing bacteria such as Prophyromonadaceae and Enterococcaceae,and reducing conditional pathogens such as Corynebacteriaceae.Chickpea also improved the metabolic function of model rats in the progression of T2DM disease.These include increased basal metabolic capacity(carbohydrate metabolism,energy metabolism),and a reduction in amino acid degradation and lipid metabolism.Thus,chickpeas can alleviate hyperglycemia through key species of the gut microbiota and restore the dysregulated microbiota composition and function to normal.