High temperature inhibits wheat grain filling. Polyamines (PAs) are closely associated with plant resistance caused by abiotic stress. However, little is known about the effect of PAs on the grain filling of wheat under heat stress. Two wheat varieties differing in heat resistance were used, and endogenous PAs levels were measured during grain filling under normal growth conditions outside the greenhouse (CK), artificially simulated high temperature (HT), artificially simulated high temperature plus exogenous application of spermine (HT + Spm) and artificially simulated high temperature plus spermidine (HT + Spd) treatments. Additionally, the variation of antioxidant enzymatic activities and osmotic adjustable substances content in grains was measured during grain filling. The results showed that compared with HT,HT + Spm and HT + Spd significantly increased grain weight of XC 6 (heat-resistant variety) by 19% and 5%, and XC 31 (heat-sensitive variety) by 31% and 34%, activity of superoxide dismutase (SOD), peroxidase (POD)and catalase (CAT) and content of Spm, Spd, and proline (Pro) increased significantly, while putrescine (Put), malondialdehyde (MDA) and soluble sugar (SS)contentdecreased during grain filling; The correlation analysis showed that grain weight was negatively correlated with the content of PUT, MDA, Pro and activity of SOD and CAT and positively correlated with the content of Spd and activity of POD in grains. Our results indicated that exogenous Spm and Spd could alleviate the heat injury of grain filling.