Abstract: OBJECTIVE To investigate the effect of puerarin on hyperglycemia in HFD mice and the role of GLP-1R signaling in modulating the hypoglycemic effect of puerarin. METHODS HFD induced diabetic mice was used as the subject， GLP-1 receptor antagonist Exendin 9-39 （Ex 9-39） was used to block the activation of GLP-1R signaling， and in vivo experiments were performed to further evaluate the role of GLP-1R signaling in the hypoglycemic effect of puerarin. RESULTS The glucose homeostasis and the OGTT (oral glucose tolerance tests) of HFD mice were both improved by puerarin， while the effect was impaired by Ex 9-39 significantly. Moreover， puerarin increased serum insulin contents in HFD mice， but such effect was blocked by Ex 9-39 as well. Elevated expression of ki67 in β-cells was observed in pancreatic sections from HFD mice pre-treated with puerarin using Ki-67/insulin immunostaining， suggesting that puerarin could promote β-cell proliferation. The puerarin effect was inhibited when the GLP-1R signaling activation was blocked by Ex9-39. CONCLUSION The results confirmed that the activation of GLP-1R signaling was critical for the anti-diabetic effects of puerarin on HFD induced diabetic mice. Our finding highlights the potential value of Radix puerariae as a dietary supplement for diabetes care.