Mechanisms Of Various Protein Sources Enteral Nutritional Formula On The Genomic Characteristics Of Gut Microbiota In Streptozotocin Induced Type2 Diabetes Mice Mellitus Mice

Diabetes Mellitus(DM)is a metabolic disease associated with the irregular metabolism of protein,lipid and glucose and it was caused by the interaction of genetic and environmental factors or target tissue cells with impaired insulin biological function.Chronic metabolic diseases of fats,water and electrolytes,and various hormonal disorders often leaded insufficiency and failure of various organs,tissues and various blood vessels.In patients with normal bowel function,nutritional therapy is a prerequisite for preventing complications and increasing infection and hospitalization rates.Enteral nutritional support is not only better than parenteral nutrition,but it can also reduce the blood sugar significantly.With reference to existing enteral nutrition suspension formulations on the market,such as Ruidai,Abbott SR and Dongze All-I,etc.,silkworm pupa protein,soy protein and whey protein were selected as the source of the protein of the enteral nutritional formulia to explore Whether the different proteins sources in enteral nutrtional formulia has different effects in adjusting the nutritional metabolism status,liver and kidney function damage and the gut microbiota change of T2 DM mice.In this experiment,silkworm pupal protein,soy protein and whey protein were used as protein sources of enteral nutritional formulia(ENF).Four different protein source ENF,Silkworm pupa protein ENF(SPP ENF),Soybean protein isolate ENF(SP ENF),Silkworm pupa protein combined with Lactalbumin protein ENF(LCSSP ENF),Lactalbumin protein ENF(LP ENF),were designed and optimized.Then supplemented with the addition of slow-release carbohydrates(fructooligosaccharides,tapioca starch and corn starch),appropriate amounts of vitamins and minerals,and 10 g / 100 g dietary fiber.These four specially designed ENF were used as experimental treatment groups,namely the soy protein group(SPG),silkworm pupae protein group(SPPG),Silkworm pupal protein binds Lactalbumin protein group(LCSSPG)and whey protein group(LG).Abbott glucerna Slow Release tablet was selected as the positive control group(PG).The normal group was CG,fed with ordinary diet,and the model group was BG which were fed with ordinary feed after the model was successfully made.A ten-week high-fat diet combined with intraperitoneal injections of streptozotocin induced type 2diabetes successfully.The body weight,fasting blood glucose(FBG)of the mice were measured every weekend and the living conditions of the mice were observed.Oral glucose tolerance test(OGTT)was performed after modeling and treatment and the area under the curve number(AUC)was calculated.After modeling,the mice in each group were given support for the corresponding ENF for five weeks.The level of total protein(TP),albumin(ALB),total cholesterol(TC),triglyceride(TG),high-density lipid,low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C)in the plasma were measured and the globulin(GLO)and the ratio of albumin to globulin ratio(ALB / GLO)were calculated.The results showed that SPG had the best hypoglycemic effect,followed by SPPG,LCSSPG and PG,and LPG was the worst.Compared with BG,the values ??of TC,TG,LDL-C in the SPG,SPPG,LCSSPG and PG decreased significantly after 5 weeks of ENF intervention.,and the values??of HDL-C increased significantly.The ratios of TP,ALB,GLO,and ALB / GLO in the SPPG and SPG groups were bring back to nearly normal levels,and there were significant differences from BG.The results showed that the SPP ENF treatment was the best one to regulate the metabolism disorders of T2DMSPPG and LCSSPG were further selected for the next study.Sixty ICR mice were randomly divided into five groups: silkworm pupal protein binding whey protein group(LCSSPG),positive control group(PG),blank group(BG),silkworm pupal protein group(SPPG)and normal group(CG).Modeling making and the ENF support was same to the previous chapter.The FBG and body weight of each group of mice were measured every week and their living conditions were observed.At the second,tenth,twelfth,and fifteenth weeks of feeding,the level of alanine aminotransferase(ALT),aspartate aminotransferase(AST),uric acid(Uric.acid),free fatty acids(FFA),and urea(Ucr)and urinary creatinine(Urea)in the plasma were measureed.Three mice were elected to collect their feces during the second week,tenth week and fifteenth week of the experiment.Samples were collected by tail-lift stimulation method,and DNA was extracted and sequenced in 16 Sr RNA V3-V4 region.The results showed that compared with BG,the ALT and AST enzyme activities in SPPG,PG and LCSSPG plasma were significantly reduced after intervention,and the concentrations of Uric.acid,Ucr,and Urea were significantly reduced,and SPPG was the closest to CG.It shows that compared with the enteral nutrition formula with low added amount of silkworm pupal protein,the formula with high added amount of silkworm pupal protein is more superior to the protection of liver and kidney function.The results of intestinal flora analysis showed that the intestinal flora of T2 DM mice was significantly different from that of normal mice.After treatment,the alpha diversity of the intestinal flora of BG mice decreased significantly.PCA principal component analysis showed that BG and CG were significantly separated and far apart.After treatment,the observed species index,chao index,ace index,shannon index,and simpson index in SPPG,PG,and LCSSPG were significantly lower than that in CG.SPPG,PG,LCSSPG are significantly separated from BG in the PCA diagram,and closer to CG.The relative abundance ofParabacteroides distasonis increased by 111.62,149.01,and 100.6 times in SPPG,PG,and LCSSPG,respectively,which was 7 times than that of CG.After treatment,the proportion of Bacteroidetes in SPPG increased significantly,and the proportion of Firmicutes and the ratio of Bacteroidetes / Bacteroides were significantly reduced(P<0.05),even lower than BG.The relative abundance of Bacteroides increased by 4.5,5.6,and 6 times in LCSSPG,PG,and SPPG,respectively.After treatment,compared with BG,the relative abundance of Prevotella in SPPG was significantly reduced.Compared with BG,the relative abundance of Clostridium and Coprococcus in LCSSPG increased significantly(P <0.05).KEGG(Kyoto Encyclopedia of Genes and Genomes)function prediction results indicated that adipose cytokine signaling pathway,ubiquitin system and phosphonate and phosphinate metabolism functions are significantly increased,phosphotransferase system(PTS)Significantly reduced,increased secondary bile acid biosynthesis in SPPG.Phosphotransferase system(PTS)function significantly increased in LCSSPG.Three ENF can significantly reduce PBG levels in T2 DM mice,and at the same time,play a role in regulating blood lipid metabolism and protein metabolism,especially SPP ENF optimizes the composition of beneficial bacteria in the intestinal tract,and selectively promotes the value-added of bacteria groups which was closely correlated with glucose metabolism,such as Parabacteroides distasonis.