Objective: To study the effect of metformin on lactate metabolism in hepatocytes in vitro under high glucose stress. Method: LO2 hepatocytes was cultured in vitro, hepatocytes were randomly divided into blank control group, 25 mmol/L glucose solution, 27 mmol/L glucose solution, 29 mmol/L glucose solution, 31 mmol/L glucose solution, 33 mmol/L glucose solution, 35 mmol/L glucose solution treatment group, after determining the optimal concentration as 31 mmol/L, use 30 mmol/L metformin solution, and then divided into blank control group, normal hepatocytes + the optimal concentration of glucose solution, normal hepatocytes + metformin solution , normal hepatocytes+. The optimal concentration of glucose solution normal hepatocytes + metformin solution, calculate the number of hepatocytes on cell count plate respectively in the 12 h, 24 h, 48 h, and use the lactic acid kit to determine the lactic acid value of the cell culture medium of normal liver cells + optimal concentration glucose solution and normal liver cells + optimal concentration glucose solution + metformin solution at 12 h, 24 h, and 48 h, respectively. Results: There was no significant change in the lactic acid concentration but significant increase in the number of surviving hepatocytes in the high-glycemic control group compared with that in the high-glycemic control group without metformin. Conclusions: Metformin has no significant effect on lactic acid metabolism of hepatocytes under high glucose stress in vitro, and has a protective effect on hepatocytes under high glucose stress. Based on this, it is preliminarily believed that metformin is not the direct factor leading to diabetic lactic acidosis.