| Objective:To explore the intestinal flora on mucosal permeability and sugar absorption mechanism for the study of diabetes control blood glucose to provide theoretical and practical basis.Methods:50 Sprague Dawley?SD?rats were selected and divided into three groups:Diabetic group?A group?;Diabetic rats with imbalanced intestinal flora group?B group?;Healthy control group/normal The group?C group?,with an average body weight of 90±5g,was fed one week after adaptation and fed with high fat and high sugar for four weeks.After fasting for 12 hours,rats with fasting blood glucose>9.0mmol/L were discarded.The remaining eligible rats were injected intraperitoneally with streptozocin?STZ?at a dose of 40 mg/kg.Fasting for 12 hours after 3 days,fasting blood glucose>16.7 mmol/L was considered to be a successful model for diabetes.One half of the rats in the diabetic group were randomly selected and daily administered with a dose of 1 mg/m L of ceftriaxone sodium.The stomach was fed once a day for one week.Fecal samples were collected after one week.After PCR-DGGE?Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis?,the number of bands in the group was significantly lower than that in the normal group.This was an imbalance model of the intestinal flora in diabetic rats.Modeling success.After successful modeling,each rat was given lactulose and mannitol by gavage at a dose of 120 mg of lactulose and 80 mg of mannitol?1 mL each?[1].Rats were given intragastric administration,each was placed in a separate sham cage,fasted for 24 hours,and urine was collected.Lactulose in urine was determined by liquid chromatography-tandem mass spectrometry?LC-MS/MS?.Mannitol output.Daily observation of the rat's appearance signs,weight and mental status were recorded.Blood glucose was measured every three days.Feces were taken every week to observe changes in intestinal flora.After euthanasia,the rat small intestine tissues were observed for morphological observation,and small intestine tissue homogenates were prepared to determine Na+-K+-ATPase in the small intestine epithelial cells.Results:1.Diabetic rat model rateIn this experiment,a total of 50 SD rats were injected with STZ at a dose of 40mg/kg to establish a diabetic model,of which 30 were modeled and the rate of molding was 60%.2.Effect of imbalance of intestinal flora on apparent signs of diabetic rats Symptoms of diarrhea in diabetic imbalance group were compared with those in diabetic group.Feces were soft and yellow in color;Diabetic rats showed slowness,apathy,unsmooth fur,severe hair loss,and body weight loss compared with normal control rats..3.Effect of imbalance of intestinal flora on body weight of diabetic ratsExperiments showed that the weight of the normal group rats was 437.8±12.69g,the body weight of the diabetic group was 233.5±6.33g,the body weight of the diabetic group was 236.2±5.20g,and the imbalance between the diabetic group and the normal group was compared.The body weight was significantly lower and the difference was extremely significant?p<0.01?.There was no significant difference between the diabetic group and the diabetic group in the imbalance group?p>0.05?.The weight of the diabetic group was significantly lower than that of the normal group.The difference was extremely significant?p<0.01?.4.Effect of imbalance of intestinal flora on blood glucose changes in diabetic ratsThe fasting blood glucose of SD rats in the normal group was 5.263±0.18mmol/L.The fasting blood glucose in the diabetic group was 23.23±1.08mmol/L.The fasting blood glucose in the diabetic group was 22.98±0.69mmol/L.Compared with the normal control group,the fasting blood glucose in the unbalanced diabetic group was significantly higher than that in the normal control group?p<0.01?.There was no statistical difference in blood glucose levels between the diabetes group and the diabetic imbalance group.Significant?p>0.05?,compared with the normal control group,the diabetes group rats fasting blood glucose was significantly higher than the normal control group?p<0.01?.5.Effect of imbalance of intestinal flora on changes of mucosal permeability in diabetic ratsExperiments showed that the ratio of lactulose to mannitol in the urine of normal rats was 0.3160±0.12.The ratio of lactulose to mannitol in urine of diabetic rats was0.7249±0.10.The urine of diabetic rats was unbalanced in urine.The ratio of fructose to mannitol was 1.541±0.33.Compared with the diabetic group,the ratio of lactulose to mannitol in the diabetic dysbalance group was higher than that in the diabetic group,with statistical significance?p<0.05?.In the unbalanced group,the ratio of lactulose to mannitol in the unbalanced group of diabetes was significantly higher than that in the normal group?p<0.01?.Compared with the normal group,the ratio of lactulose/mannitol increased in the diabetic group compared with the normal group.Statistically significant?p<0.05?.6.Intestinal flora imbalance in diabetic ratsThe experiment showed that the number of bands in the normal control group was30.00±1.47,the number of bands in the diabetic group was 22.00±0.58,and the number of bands in the diabetic imbalance group was 9.667±0.33.The imbalance between the diabetic group and the diabetes group was compared.The number of bands decreased,which was statistically significant?p<0.01?.Compared with the normal control group,the number of bands decreased in the diabetic group and there was a statistically significant difference?p<0.01?.The diabetic group was compared with the normal control group.In comparison,the number of bands decreased,which was statistically significant?p<0.01?.7.The effect of imbalance of intestinal flora on the morphology of small intestine in diabetic ratsAfter observing the small intestine histomorphology in the duodenum,jejunum and ileum of 3 rats in each group,it was found that the duodenal muscle layer of the diabetic imbalance group was thinner than the normal group,and the jejunal villus swollen.Intestinal and ileal villus breakage phenomenon is obvious,ileum submucosal lymphocyte infiltration.In the diabetic group,the villi were arranged irregularly,and the fracture was serious.The ileum segment was particularly prominent.The jejunum and ileum muscle layers had obvious holes,and there was no obvious change in the normal control group.8.Effects of imbalance of intestinal flora on changes of Na+-K+-ATPase in intestinal epithelial cells of diabetic ratsExperiments showed that the content of Na+-K+-ATPase in the small intestine epithelial cells of normal rats was 2.812±0.25ng/mL,and the content of Na+-K+-ATPase in the small intestine epithelial cells of diabetic rats was 1.877±0.13ng/mL.The content of Na+-K+-ATPase in the intestinal epithelial cells of the unbalanced group was 1.500±0.07ng/mL.Compared with the diabetic group,theNa+-K+-ATPase content in the intestinal epithelium of the diabetic group was lowerthan that in the diabetic group.Significance?p<0.05?.The content of Na+-K+-ATPasein rat intestinal epithelial cells was significantly lower in the diabetic group and inthe normal group than in the unbalanced group?P<0.01?.The difference wasstatistically significant?p<0.01?.Compared with the normal group,the content ofNa+-K+-ATPase in the small intestine epithelial cells was significantly decreased,which was statistically significant?p<0.01?.Conclusion:1.A rat model of type 2 diabetes was successfully established at a dose of 40 mg/kg streptozotocin?STZ?.2.Low doses of ceftriaxone sodium in diabetic rats once a day,continuous gavage for a week can cause imbalance of intestinal flora in diabetic rats.3.The activity of Na+-K+-ATPase in intestinal epithelial cells of diabetic imbalanced rats was significantly decreased.4.Diabetic bacterial imbalance in rat intestinal barrier permeability. |
PDF Full Text Request