| Objective:1.To insure target gut microbiota by determine the level of Veillonella spp and C.coccoides subgroup in fecal total microbiota DNA of Normal Glucose tolerance（NGT）and Type 2 Diabetes Mellitus（T2DM）in Kazak and Uygur in Xinjiang;2.To study the relationship of target gut microbiota and metabolism profiles of plasma amino acid and type 2 diabetes.Methods:1.Compared the level of Veillonella spp and C.coccoides subgroup in fecal total microbiota DNA of NGT and T2DM in Kazak and Uygur in Xinjiang by using 16SrDNA real time PCR;2.Male C57BL/6 mice were divided into four groups:common chow diet-fed normal animals（Normal）,berberine（200mg/kg）-treated common chow diet-fed animals;high fat diet and 20%high fructose corn syrup（HFCS）for drinking animals（HFD+HFCS）;berberine（200mg/kg）-treated HFD+HFCS animals.Identical volumes of distilled water were given to normal and HFD+HFCS animals.At 4、8、12、16weeks,random drawing 12 mouse in each group,detecting the level of fasting glucose（FBG）,random blood glucose（GLU）,total cholesterol（TC）,Triglyceride（TG）,Fasting serum insulin（FINS）by the blood of tip of the tail,and calculate Insulin resistance index（HOMA-IR）,Insulin sensitive index（ISI）,after Oral glucose tolerance test（OGTT）and Insulin tolerance test（ITT）,the animal were weighed and anesthetized by intraperitoneal injection of pentobarbital collecting blood and colon contents;3.Determine the level of target gut microbiota at each time point by 16SrDNA real time PCR;4.Use HPLC to determine the plasma amino acid profile of mice.Reslut:1.16SrDNA real time PCR showed in genus level,the abundance of Veillonella spp and C.coccoides subgroup in Kazak T2DM group were much higher than Kazak NGT group（p<0.05）;2.Compared with Normal mice,the contents of GLU、FINS、TC were significantly increased（p<0.05）in HFD+HFCS mice at 4 weeks.At 8 weeks,Compared with Normal mice,HFD+HFCS mice appear impaired glucose tolerance,the AUC of glucose in ITT and HOMA-IR were significantly increased（p<0.05）,ISI was significantly reduced.The contents of FBG in HFD+HFCS mice became much higher than normal mice at 16 weeks.berberine significantly lower TC、FINS、HOMA-IR,and increase ISI,and Improve impaired glucose tolerance and insulin resistance in HFD+HFCS mice;3.Failed to amplify target fragments of Veillonella spp in colon contents total microbiota DNA of mice;Compared with normal mice,the level of Desulfovibrio was significantly increased,C.coccoides subgroup was significantly decreased since 4 weeks,berberine significantly decreased Desulfovibrio,and decrease C.coccoides subgroup,but have no effect on Desulfovibrio and C.coccoides subgroup in normal mice;4.Sice 8 weeks,Compared with the normal mice,the metabolism profiles of amino acid in the HFD+HFCS mice were significantly different,berberine significantly inhibit the change of the metabolism profiles of plasma amino acid induced by high fat and HFCS.Pattern recognition method was applied 6 amino acid markers were different between all the groups（Asp,Thr,Gly,Cys,Tyr,Lys）;5.The result of Path analysis shows modeling or not,the content of Desulfovibrio,Asp,Lys,Thr and Glu could positive impact the content of FINS Directly,modeling or not could indirect effects the content of FINS by impact on Desulfovibrio,Asp,Lys and Thr.Besides,Desulfovibrio also could indirect effects the content of FINS by impact on Asp,Lys and Thr.Concludes:1.The Differences trend of Veillonella spp and C.coccoides subgroup are different between human and mice,the relationship between these two gut microbiota and type 2 diabetes remains to be further studied;2.Intestinal flora may be able to affect the levels of plasma amino acids,which affects the body glucose and lipid metabolism. |
