Nowadays diabetes, as a metabolic disorder, is increasing at an alarming rate. Glycation and production of advanced glycation end products (AGEs) is the most important factor involved in diabetic complications. Due to the side effects of synthetic drugs, the demand for natural anti-diabetic herbal medicines has increased. Propolis is a natural and resinous material, which iscollected by honeybees. Due to the impact of nanotechnology in medicine and the advantageous role of nanoparticles in treatment, nano-propolis particles (PNP) were prepared. The anti-glycation effect of PNP at various concentrations was investigated on human hemoglobin (Hb) glycation and fructation and compared with aspirin as a common anti-glycation agent using glycation specific AGE fluorescence, AGE-specific absorbance and circular dichroism (CD) methods. Fluorescence spectroscopy results showed that PNP inhibited the formation of AGEs in Hb glycation and fructation by glucose and fructose, respectively. CD results revealed that PNP caused an increase in Hb beta-sheet content while decreasing the alpha helical content. Additionally, the results of UV-Vis spectroscopy and fluorescence emission of heme degradation products revealed the protective effect of PNP on heme during glycation and fructation of human Hb. It is notable that the synergistic effects of combined propolis nanoparticles and aspirin is more than either of them alone. However, having said that, PNP as a natural product has a potential to be an effective drug in the treatment of diabetes.