Research On Prevention Of Type 2 Diabetes Mellitus By Regulating The Intestinal Flora Disorder Of ZDF Rats By Salsalate

Objective: According to the latest guidline,the prevention of diabetes should be early.Clinical studies have found that salsalate can regulate metabolic disorders of glucose and lipid,but the mechanism is not clear.A large number of studies have found that intestinal flora disturbance is closely related to T2 DM.So we selet ZDF rats as the study object.Salsalate is given in IGT stage(50mg/kg.d-1),observe its improvement of blood lipid,glucose and intestinal flora disorders,preliminarily explore the mechanism of preventing type 2 diabetes of salsalate.Method: ZDF rats as model ZL rats as control are tested OGTT every week.Based on the blood levels,the age of ZDF rats are divided into three stages: NGT,IGT and DM stage.Select 10 of ZDF rats to feed with salsate in IGT stage.Lipid index are detected by biochemical method.Detect insulin and glucagonlevel,liver and kidney function with biochemical method.Use 16 s r DNA sequencing technology to test the gene of intestinal flora.Use light microscope and electron microscope to observe shape and structure of ileum.Use immunohistochemical method to detect the ileum junction protein(ZO-1,Occludin).Use ELISA to measure LPS level of intestinal tissue solution.Use immunofluorescence method to test the expression of M1 and M2 macrophages.Use RT-PCR method to detect the macrophage related m RNA(IL-1,IL-4,IL-10,IL-12,i NOS,Arg-1)levels.Use immunohistochemical method to detect the inflammatory factors(TIL4,NFкB,TNFα).Results: A,intervention effect of salsalate of improving glucose and lipid.1.At 9-week-old,all of the ZDF rats can be diagnosed as IGT.At 12 week-old,all the model group rats can be diagnosed as DM.Only one of intervention group developed IGT,others reversed to NGT.2.In the whole progression,AUCg of ZDF rats was higher than ZL rats(P﹤0.05).At 12 week-old,AUCg of the model group is larger than intervention group(P﹤0.05)and it showed that TG,TC,HDL of intervention group are lower than model group.3.With the progression of diabetes,insulin secretion in ZDF rats was gradually decreased,but it’s relatively constant in ZL group.Insulin secretion is increased and glucagon is decreased in intervention group than model group(P < 0.05).HOMA-IR is lower of intervention group than model group.4.At the end of the experiment,there are no significant difference among the three groups in the liver and kidney function and 24 hours urine protein.B,the effect of salsalate on intestinal microflora in ZDF rats and the mechanism of preventing diabetes.1.The alpha diversity is decreased with the progression of diabetes in model group,but the ZL rats is relatively constant.The alpha diversity of intestinal microflora is higher in intenvention group than model group.Beta diversity is more closed to normal group than model group in intervention group.2.The relative abundance of actinobacteria of normal group is higher than model group in NGT stage.With the progress of disease,the relative abundance of firmicutes,proteobacteria,verrucomicrobia are decreased,and the relative abundance of bacteridetes is increased in model group.Relative abundance of firmicutes and verrucomicrobia.has no significant difference between intervention group and normal group.3.In NGT stage the relative abundance of bacteroides,lachnospira,ruminococcus,roseburia of normal group is lower than model group,and the relative abundance of lactobacillus,bifidobacterium is higher in normal group.The relative abundance of ruminococcus,clostridium,lachnospira,roseburia,sutterella,akkermansia are gradually decreased,and the relative abundance of prevotella is inceased in model group during progression.The relative abundance of roseburia is higher in SAL group than model group.4.Microscope shows that ZDF rats intestinal epithelial integrity,tight junction are getting worse and junction protein ZO-1 and Occludin are decreased than normal and intervention group.5.The level of LPS of ileum is increasing with the progression of diabetes,M1 macrophages and markers’ level are higher and M2 macrophages and markers’ level are lower in model group compared with normal and intervention group.Conclusion: 1.Low dose of salsalate early intervented can improve glucose and lipid metabolism,promote the secretion of insulin,reduce glucagon secretion,improve insulin resistance.2.Salsalate can regulate the intestinal flora disorder and reduce the production of LPS,protect the intestinal structure and function,prevent LPS into the blood,reduce macrophage polarization,thereby reduce the production of inflammatory cytokines,promote the formation of anti-inflammation factor,improve the state of chronic inflammation in ZDF rats,to prevent the progression of T2 DM.3.Low dose of salsalate in prevention of T2 DM is safe.