Oxidative stress is a promoting factor in the pathologic process of glucocorticoid - induced osteoporosis (GIO), while the mechanism is still unclear. Thioredoxin-interacting protein (TXNIP) is a vital protein responsible for regulation of cellular reactive oxygen species (ROS) generation elicited by mitochondrial oxidative stress, and which may activate oxidative phosphorylation under the pathogenic status. In this research, the results showed that signaling pathway associated with the mitochondrial oxidative phosphorylation (MOP) down-regulated under conditions of TXNIP siRNA in MG63 cells. Furthermore, the evidence revealed that the expression level of TXNIP in serum and bone was elevated in a rat of GIO. Moreover, the differential proteins (Ndufs3, SDHD, Cyt B, COX IV, and ATP B) related to MOP pathway were identified to down-regulate in the proteomics of bone tissues by using isobaric Tags for Relative and Absolute Quantification (iTRAQ) method in TXNIP knockout mice treated with glucocorticoid, and the proteins were also verified by simple western blot. Taken together, the present findings highlights that TXNIP involves in triggering the process of bone loss via up-regulation of the MOP pathway, resulting to GIO, while TXNIP knockout can prevent the pathogenesis of GIO to some extent.