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Abstract Type 2 diabetes is a complex metabolic and endocrine disorder worldwide, which causes severe health and economic problems. The effects of polysaccharide from Plantago asiatica L. (PLP) on high-fat diet and streptozotocin-induced type 2 diabetic rats were examined. Administration of PLP caused significant decreases in the concentrations of blood glucose, insulin, total cholesterol, triglyceride, non-esterified fatty acid and maleic dialdehyde, and significant increases in the levels of high density lipoprotein-cholesterol and the activities of antioxidant enzymes compared with diabetic rats after 4 weeks’ treatment. The concentrations of short-chain fatty acids (SCFA) were significantly higher in the feces of diabetic rats after treatment with PLP. Moreover, colon bacterial diversity and abundance of bacteria, including Bacteroides vulgatus, Lactobacillus fermentum , Prevotella loescheii and Bacteroides vulgates were significantly increased by PLP treatment. These results indicated that the anti-diabetic effect of PLP in type 2 diabetic rats may be associated with regulation of gut microbiota and increased levels of SCFA. Graphical abstract Image Highlights • PLP attenuated hyperglycemia and hyperlipidemia in Type 2 diabetic rats. • PLP administration suppressed oxidative stress in Type 2 diabetic rats. • SCFA was significantly increased after PLP intake in Type 2 diabetic rats. • PLP improved structure of colon microbiota in Type 2 diabetic rats. [ABSTRACT FROM AUTHOR]