Global cerebral ischemia and reperfusion injury (GCI/R) can lead to neuronal apoptosis and contributes to permanent neurological sequelae. However, the underlying mechanism is largely unknown. Therefore, the present study aimed to assess the effects of GCI/R on the tribbles homolog 3 (TRB3) and to explore the role of TRB3 in GCI/R. The GCI/R model was developed in Sprague-Dawley male rats by four-vessel occlusion. Subsequently, the expressions of TRB3, endoplasmic reticulum stress markers, and apoptosis-associated proteins were examined by western blot at 1 h, 6 h, 12 h, 24 h, and 72 h after GCI/R. TRB3 short-hairpin RNA (shRNA) lentivirus was constructed and used to investigate the role of TRB3 in GCI/R-induced neuronal apoptosis. GCI/R increased the level of TRB3, endoplasmic reticulum stress markers, and pro-apoptotic proteins. The level of protein kinase B (Akt) phosphorylation was reduced during GCI/R. Administration of TRB3 shRNA lentivirus attenuated GCI/R-induced up-regulation of TRB3, endoplasmic reticulum stress, and neuronal apoptosis. Furthermore, TRB3 shRNA lentivirus reversed the reduced level of Akt phosphorylation induced by GCI/R. These data implied that TRB3 participated in the GCI/R-induced neuronal apoptosis. Knocking down TRB3 attenuated endoplasmic reticulum stress, enhanced Akt phosphorylation, and protected neurons from apoptosis in response to GCI/R. These results demonstrated that the downregulation of TRB3 may be a promising approach for treating GCI/R. (C) 2017 IBRO. Published by Elsevier Ltd. All rights reserved.