〔Abstract〕 Objective To investigate the effects of different concentrations of glucose on the energy metabolism of red blood cells(RBC) in vitro and its molecular mechanism. Method Fresh anticoagulant blood was collected from 6 healthy adults. The white blood cells and plasma were removed by centrifugation. After washing RBC three times with PBS respectively, the 6 people's RBC were gently mixed and divided into 4 groups. D-glucose was added to make a final concentration of 0, 6, 20, 30 mmol/L, respectively. After incubation at 37 ℃ for 0, 24 and 48 hours, some of the RBC suspension was taken out to detect eryptosis, ATP concentration, hexokinase(HK) activity, pyruvate kinase(PK) activity, reactive oxygen species(ROS) content and hemolysis rate by using flow cytometry and multi-function microplate reader. The experimental results were analyzed by one-way ANOVA. Results Compared with the 0 mmol/L group, after incubating RBC with glucose at 6, 20, 30 mmol/L for 24 h and 48 h,the HK activity,PK activity,and ATP content in the RBC all increased,and upward trend with increasing glucose concentration,and the difference was statistically significant( P<0. 05); while ROS content and decay rate decreased,and decreased with increasing glucose concentration,the difference was statistically significant( P<0. 05). After culturing erythrocytes with different concentrations of glucose( concentrations of 0,6,20,30 mmol/L) for 48 h,the ROS content and erythrocyte decay rate in RBC of different concentrations increased with the incubation time,compared with 0 h,the difference was statistically significant( P<0. 05); HK activity and PK activity gradually decreased,compared with 0 h,the difference was statistically significant( P<0. 05). The ATP content decreased in a fluctuating manner,and compared with 0 h,the difference was statistically significant( P<0. 05). Conclusion Short-term high-glucose culture of RBC in vitro not only strengthens the energy metabolism in RBC,but also reduces the oxidative stress and eryptosis. However,no glucose supply leads to gradually exhausted ATP and increased oxidative stress and eryptosis.