Effects Of Milk-borne Composit Probiotic On Glucagon-Like Peptide-1 In Db/db Mice

Objective:To investigate the anti-diabetic effect of milky source compound probiotic milk on db/db diabetic mice.The compound probiotics contain fourteen kinds of probiotics such as Lactobacillus helveticus,Lactobacillus plantarum,Lactobacillus paracasei,Lactobacillus paracasei subsp.tolerans,Lactobacillus kefiri,Lactobacillus harbinensis,Lactobacillus hilgardii,Lactobacillus pentosus,Lactobacillus kefianofaciens,Lactococcus lactis,Kluyveromyces marxianus,Pichia membranifaciens,Candida ethanolica,Issatchenkia orientalis.And its effect on the secretion of glucagon like peptide GLP-1 and its related gene expression in db/db diabetic mice.Methods:1.Fourteen probiotics freeze-dried powder were activated and cultured separately.The activity of probiotics in aseptic defatted camel milk was determined by the method of resazurin.And the drug resistance of 8 strains of probiotics was tested by Agar diffusion method(K-B method).In addition,preparing 14 kinds of probiotic milk from milk source in laboratory.2.50sPF grade 6-week-old male db/db diabetic mice diabetic mice were divided into five groups and treated for 6 weeks with Positive drug group I(oral 0.3 g/kg metformin hydrochloride),positive drug group 2(subcutaneous injectionliraglutide of 0.2 ug-g),low-dose milk of probiotic probiotics 1.0×108 lactic acid bacteria 1.0×10 6 of Eysa oriental yeast,and high dose of milk reproducing.1.0×10 10 lactic acid bacteria and 1.0×10 8 yeast in probiotic milk group.In addition,10male C57BL/Ks mice of the same age were used for comparison with the same dose of normal saline for 6 weeks.Fasting blood glucose(FBG)and body weight(BW)were measured before treatment and every week thereafter.All mice received OGTT in the end of 6 weeks to evaluate it glueose metabolism and to detect C-peptide in serum of db/db diabetic mice by enzyme linked immunosorbent assay(Elisa).The effects of TC,TG,LDL-C)and HDL-C)and HbAlc in plasma,liver index,renal index and insulin resistance index were calculated to observe the pancreatic tissue and liver tissue.The effect of renal histopathological detection;urine collection using an automatic biochemical analyzer to detect 24h ALB,urine sugar.3.The content of glucagon like peptide 1(GLP-1)in serum of db/db diabetic mice was detected by Elisa;The expression of Glucagon like Peptide receptor(GLP-1R),Proglucagon Gene(GCG)and Prohormone invertase(PC3)in db/db diabetic mice intestine by Real-time fluorescence quantitative PCR.Results:1.Lactobacillus helveticus,Lactobacillus plantarum,Lactobacillus kefianofaciens,Lactococcus lactis,Lactobacillus paracasei,Lactobacillus paracasei subsp.tolerans,Lactobacillus harbinensis,Lactobacillus hilgardii 8 strains of lactic acid bacteria were resistant to vancomycin.However,which were sensitive to gentamicin,amoxicillin,doxycycline,ciprofloxacin,azithromycin,ceftazidime,cefmendo respectively.2.After 6 weeks of drug intervention,High dose milk source compound probiotic milk decreased FBG(P<0.05),OGTT(P<0.05),HbA1c(P<0.05)and TG(P<0.05),TC(P<0.05),LDL-C(P<0.05),increased C-peptide(P<0.05)levels.In the high dose group db/db diabetic mice,the structure of pancreatic microlobes was intact in HE staining.,increasing of number of islet cells.And the liver tissue was stained with HE staining showed that milky source compound probiotic milk could significantly reduce the hepatic fat and edema in db/db diabetic mice,and significantly reduce the liver index of db/db diabetic mice(P<0.05).3.After 6 weeks of drug intervention,In the high dose group db/db diabetic mice the serum GLP-1 content was significantly increased(P<0.05),and the expression of GLP-1R(P<0.05)and GCG(P<0.001)were significantly up-regulated in the probiotic milk group.Conclusion:The results of this study indicates that the milky source compound probiotics milk decreased FBG,OGTT and HbA1c,increased C-peptide,modulated lipid metabolism and protected liver and kidney injury in db/db diabetic mice,which may be significantly effect on the secretion of glucagon like peptide GLP-1 and its related gene expression in db/db diabetic mice.