This study was designed to investigate the hypoglycemic effect of Agaricus blazei Murrill extracts and its beneficial effects on gut microbiota in diabetic rats. The diabetic rats were treated with ethanol and ethyl acetate extract of Agaricus blazei Murrill for four weeks. The samples of blood， liver tissues and feces were collected to evaluate the level of fasting blood glucose and Ⅱlumina HiSeq platform was used for high throughput sequencing. Results showed that supplementation with Agaricus blazei Murrill extracts could decrease the urine output and improve the mental state in diabetic rats. The symptoms of polydipsia， polyuria， polyphagia， and weight loss in diabetic rats were alleviated. Agaricus blazei Murrill extracts could cause reduction of the level of fasting blood glucose. Specifically， the ethyl acetate extract could significantly decrease the level of fasting blood glucose in diabetic rats （P<0.05）. The Agaricus blazei Murrill treated rats caused the up-regulation of Bacteroidetes and down-regulation of Firmicutes. The abundance and diversity of gut microbiota in diabetic rats were improved. The study suggested that Agaricus blazei Murrill extracts may play a role in the treatment of diabetes through regulating the structure of gut microbial.