| Background and Aims:Diabetes is a group of metabolic diseases characterized by hyperglycemia,with an increasing number of patients,which has exerted a huge impact on the medical care and social and economic development of human beings.Up to now,there are no Diabetes treatment drugs with significant curative effect and no side effects,This paper studied the therapeutic effect of intestinal probiotics combination that can directly consume glucose in the intestinal tract on diabetic mice,and explored its relevant mechanism in the treatment process,providing a new idea for the treatment and prevention of diabetes.Intestinal probiotics use glucose in the intestinal tract to accelerate the metabolism of glucose in the human body.The glucose content in diabetic patients is high.Can the participation of intestinal probiotics reduce the concentration of glucose in the body?In order to verify this effect and explore the relevant mechanism,Lactobacilli MRS Broth(MRS)was selected to screen 8 strains of probiotics in the laboratory,and the sugar consumption capacity of each probiotics was evaluated by measuring the residual glucose content in the medium.Five strains of probiotics with high sugar consumption capacity were selected to test their probiotics,including acid resistance,bile salt resistance,bacteriostasis,cell adhesion and inhibition of pathogenic bacteria adhesion.In animal experiments,Streptozocin(STZ)induced diabetes mouse model and sugar-water feeding induced diabetes mouse model were selected,and 5 probiotics combinations were selected for treatment.The results showed that the combination of probiotics had a good therapeutic effect on both types of diabetes model mice.Method one:STZ induced diabetic mouse model1.Screening of hypoglycemic strains.Choose 8 strains of probiotics activation(Lactobacillus bulgaricus,L.rhamnosus L12,L.acidophilus,L.plantarum HM218749,Bifidobacterium.animalis subsp.lactis LPL-RH,B.longum subsp.longum BAMA-B05/BAu-B 1024,L.gasseri,Streptococcus thermophilus),according to 1%of the amount of inoculated in MRS,respectively at 0,4,8,12,16,20,24 h testing residual glucose content in culture medium,to choose the hypoglycemic high ability of probiotics.2.Detection of probiotics.The selected probiotics were tested for their probiotics,including acid resistance test,bile salt resistance test,antioxidant test,bacteriostatic test,cell adhesion test and pathogen inhibition test.3.Establishment of diabetic mouse model induced by STZ.Eight-week-old male kunming mice were divided into normal control group(C group),model group(S group),STZ+sugar-water group(SG group)and probiotic combination treatment group(SGP group),with 8 mice in each group.Mice diabetes model was constructed by intraperitoneal injection of STZ(40 mg/kg/day)for 5 consecutive days.4.Treatment of diabetic mouse model.After successful modeling,SGP group was given probiotic combination by gavage every day for a total duration of 7 weeks.5.Therapeutic effect and mechanism of probiotics combination on stz-induced diabetic mice.Blood glucose and body weight of each group of mice were measured weekly to evaluate the therapeutic effect of probiotic combination on diabetic mice.When it was found that the blood glucose of diabetic mice was significantly improved after treatment,the glucose tolerance test was conducted to observe the body’s ability to regulate glucose.Western blot was used to detect the expression of proteins in the pancreatic tissue inflammatory signaling pathway(TLR-4,NF-κB/p-NF-κB)and barrier proteins in the small intestine(ZO-1,Claudin 1).The expressions of glucose transporters(SGLT-1,GLUT-2)and inflammatory cytokines(TNF-α,IL-1β,IL-6)and PDX-1 in pancreatic tissues were detected by q-PCR.The levels of insulin in each group were analyzed by immunofluorescence section of the pancreas.Method two:mice were fed with sugar water1.Establishment of sugar water feeding mouse model.Eight-week-old male kunming mice were divided into normal control group(group C),model group(group G)and probiotic treatment group(GP),with 8 mice in each group.Mice were fed 5%sugar water every day to construct the hyperglycemic mouse model.2.Treatment of mouse model fed with sugar water.Mice in the GP group were given a combination of 109 CFU/100 μL probiotics by gavage every day.3.Therapeutic effect and mechanism of probiotics combination on sugar-water fed mice.Blood glucose,fecal blood glucose and body weight of each group of mice were detected weekly to evaluate the therapeutic effect of probiotic combination on diabetic mice.The ability of mouse pancreas to secrete insulin was evaluated by immunofluorescence of pancreatic tissue sections in each group.Glucose tolerance test was used to evaluate the ability of mouse body to regulate glucose concentration.Western blot was used to detect the expression of proteins in the inflammatory signaling pathway(TLR-4,NF-κB/p-NF-κB)and intestinal barrier proteins(ZO-1,Claudin 1)in the small intestine.q-PCR was used to detect the expression of inflammatory cytokines(TNF-α,IL-1β,IL-6),PDX-1,and glucose transporters(SGLT-1,GLUT-2)in pancreatic tissues at the transcriptional level.Result one:STZ induced diabetic mouse model1.The results of in vitro hypoglycemic experiment of probiotics showed that L.thamnosus L12,L.acidophilus,L.plantarum HM218749,Bifidobacterium.animalis subsp.lactis LPL-RH,and B.longum subsp.longum BAMA-B05/BAu-B1024 had the strongest glucose consumption capacity.2.In vitro probiotics experimental results showed that all the five probiotics had good acid resistance,bile salt resistance,bacteriostasis,adhesion to small intestinal cells and inhibition of pathogenic bacteria adhesion to cells.3.The results of blood glucose and fecal blood glucose in mice showed that the blood glucose level of SG group was the highest,while that of SGP group was inhibited.4.The results of glucose tolerance test showed that after several weeks of probiotics treatment,SGP group mice could regulate their glucose level in the same time.5.Western blot results showed the SGP group of mice that probiotics treatment lowered the inflammation of the pancreas tissue signaling pathways(TLR-4,NF-κB/p-NF-κB)protein expression,and increased intestinal barrier(ZO-1,Claudin 1)expression.In the gene transcription level(q-PCR)suppresses pancreatic tissue proinflammatory factor(TNF-α,IL-1β,IL-6)expression,and raised the PDX-1 expression.Mice intestinal glucose transporters(SGLT-1,GLUT-2)expression were inhibited;6.The results of immunofluorescence sections of the pancreatic tissue showed that after STZ injection,the pancreatic tissue of the mice was severely damaged,and the secretion of insulin was greatly reduced,while the secretion of insulin was increased in the probiotic treatment group(SGP group).Result two:the mouse model fed with sugar water1.The results of blood glucose and fecal blood glucose in mice showed that the blood glucose level of mice fed with sugar water increased,while the blood glucose and fecal blood glucose levels of mice in the probiotic treatment group(GP group)decreased.2.The results of glucose tolerance showed that the ability to regulate glucose concentration was enhanced in the GP group.3.Western blot results showed the probiotic treatment group called GP group reduced inflammation of the pancreas tissue signaling pathways(TLR-4,NF-κB/p-NF-κB)protein and increased the intestinal tissue barrier(ZO-1,Claudin 1)protein expression.In the gene transcription level(q-PCR)suppresses pancreatic tissue proinflammatory factor(TNF-α,IL-1β,IL-6)expression,and increased the expression of the PDX-1 expression,at the same time,the intestinal transporters(SGLT-1,GLUT-2)expression were suppressed;4.Immunofluorescence results of pancreatic tissue sections showed that insulin in the model group decreased,while insulin in the treatment group increased to the normal level.Conclusion:1.Our in vitro hypoglycemic experiment proved that probiotics can consume a large amount of glucose;2.The results of in vivo animal experiments prove that probiotics can eventually reduce blood glucose after entering the intestinal tract;3.In addition to the major hypoglycemic effect,probiotics have been found to improve diabetes through several adjuvant pathways in vivo.Probiotics inhibit inflammation in diabetic mice by regulating the pancreatic TLR-4-NF-κB pathway and the expression of pro-inflammatory factors.Probiotics also improved intestinal barrier function in diabetic mice.4.We studied the direct consumption of glucose by probiotics and its improvement in the treatment of diabetes,indicating that the combination of probiotics is expected to be developed into food,health care products and drugs for the adjuvant treatment of diabetes. |
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