The invention relates to a probiotic composition, and discloses a probiotic composition beneficial for a glycolipid metabolic function and a preparation and application of the composition. The probiotic composition comprises some of the following nine probiotics: bifidobacterium longum, bifidobacterium breve, lactobacillus gasseri, lactobacillus rhamnosus, lactobacillus salivarius, lactobacillus crispatus, lactobacillus plantarum, lactobacillus fermentum and lactobacillus casei. Different strains in the probiotic composition cooperate with each other, so that the probiotic composition has thefunction of adjusting glycolipid metabolism, and has certain effect on serum total cholesterol, triglycerides, insulin beta-cell functions and diabetes mellitus improvement.