Objective Glycer-aldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme, has long been considered a housekeeping enzyme.It has been researched as an important role of glycolytic those thirty years.Recently some studies show that administration of some stimuli could induce GAPDH S-nitrosylation and facilitate its combination of Siahl, the latter of which could induce GAPDH nucleus translocation and then cell apoptosis.The objective of this study is primarily to determine whether GAPDH could be S-nitrosylated, then boost the combination of GAPDH and Siahl induced its nucleus translocation and cell death via GAPDH S-nitrosylation.Methods Ischemic model is rat four-vessel occlusion (4-VO) ischemic model.S-nitrosylation was examined mainly by biotin switch assay.Protein expression and protein interaction were determined by immunoblotting and immunoprecipitation respectively.Results Here we show that S-nitrosylation of GAPDH was induced by cerebral ischemia-reperfusion.In cytoplasm, administration of MK801, an antagonist of N-methyl-D-aspartate receptor (NMDAR), or 7NI, inhibitor of nNOS, or DEP, the monoamine oxidase-B (MAO-B) inhibitor, diminished the induced S-nitrosylation of GAPDH and also the combination of GAPDH and Siahl.Conclusion In nucleus, GAPDH S-nitrosylation facilitates its nuclear translocation.Administration of the 3 drugs can diminish the nuclear translocation of GAPDH and also diminish the combination of GAPDH and Siah1.