Objective: To study the effect of probiotics on gut microbiota in Type 2 diabetes patients and its clinical application value. Methods: Select Type 2 diabetes patients to take orally probiotics for 24 weeks, collect stool samples of subjects at the baseline and end of the trial, identify and analyze gut microbiota of each sample by 16srRNA high-throughput sequencing, and compare the changes of blood glucose, blood lipid and insulin resistance before and after the intervention. Results: A total of 75 patients completed clinical observations. 16srRNA high-throughput sequencing showed that the proportion of the subjects with increased Actinobacteria and Tenericutes at the end of the trial has increased (37.8% and 75.7% respectively). The genus level analysis showed that the number of subjects with increased intestinal probiotics and with decreased conditioned pathogens all increased. Cluster analysis before and after intervention showed that the gut microbiota of samples in the same group had a higher similarity. Compared with the subjects at the baseline status, at the end of the trial after the intervention, fasting blood glucose (FBG) of the subjects significantly decreased (P<0.05), the proportion of the subjects with triglyceride (TG) and cholesterol up to standard increased, and HOMA-IR was significantly improved (P<0.05). Conclusions: Probiotics can regulate the gut microbiota of Type 2 diabetes patients, promote fasting blood glucose (FBG) to reach the standard and improve insulin resistance, and help improve lipid metabolism