Objective To investigate the effects of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) gene expression on the formation of extracellular trap (NET) and glycolytic metabolism in chronic obstructive pulmonary disease (COPD). Methods Bioinformatics was used to explore the correlation between MTHFD2 and glycolysis and neutrophil (NEU) markers. Subsequently, extracellular DNA content was detected by Picogreen staining assay. The expression levels of neutrophil extracellular trap (NET) related markers MPO and cit-H3 were detected by immunofluorescence staining. The expression levels of MTHFD2 and glycolytic related genes (HK1, PDK1, SLC2A1 and MYC) were detected by qRT-PCR. NET formation levels, glucose and lactate contents in NC neutrophils, neutrophils overexpressing MTHFD2, and neutrophils overexpressing MTHFD2+ glycolysis inhibitor (2-DG) were detected by cellular and molecular experiments, respectively. Results The results of bioinformatics showed that MTHFD2 was highly expressed in the tissues of COPD patients, and was positively correlated with genes related to glycolysis pathway and markers of neutrophils. Immunofluorescence staining and Sytox Green staining showed that the level of NET in COPD was significantly increased. The results of cell and molecular experiments showed that overexpression of MTHFE2 could induce the expression of HK1, PDK1, SLC2A1, MYC, promote the decomposition of glucose into lactic acid, and then promote the formation of neutrophil NET. Conclusion This study explores the effect of MTHFD2 gene on the formation of neutrophil NET and glycolysis metabolism, analyzes the relevant mechanism of MTHFD2 promoting the formation of NET in COPD patients through glycolysis, and provides new potential diagnostic markers and therapeutic targets for the treatment of COPD patients