The objectives of this research were to extract and purify two water soluble polysaccharides from Borage leaves, further explore the structural characteristics and evaluate the hypoglycemic, anticancer and immunological activities. Two water-soluble polysaccharides were extracted and purified from Borage leaves by cellulose enzyme assisted extraction (BLP-1) and microwave assisted extraction (BLP-2), respectively. The physicochemical properties of BLP-1 and BLP-2 were investigated and compared. The hypoglycemic activities of BLP-1 and BLP-2 were evaluated by detecting the inhibition rate of α-glucosidase and α-amylase, the anticancer activities were evaluated by detecting the inhibition rate of HepG2, MX-1, and A549 cells, and the immune activities were evaluated by detecting the secretion of the immune factors of RAW246.7 cells. Results showed that BLP-1 and BLP-2 had different sugar, and protein contents. BLP-1 and BLP-2 were consisted of Ara, Gal, Glc, Xyl, and with molecular weights (Mws) of 20.1 and 22.6 kDa, respectively. Thermal gravimetric analysis (TGA) suggested that BLP-2 had more a stable structure than BLP-1. The two polysaccharides have potential for inhibiting α-glucosidase and α-amylase activities, suppressing HepG-2, A549 and MX-1 cancer cells proliferation, and activating macrophage RAW 264.7 cells to secret immune cytokines for mediating cellular immune response. BLP-1 and BLP-2 could be used as a bioactive ingredient in new functional foods and pharmaceutical products,which provided a theoretical basis for further study on the structure-activity relationship.