Adriamycin (ADR)-induced nephropathy in animals is an experimental analogue of human focal segmental glomerulosclerosis (FSGS), which presents as severe podocyte injury and massive proteinuria and has a poorly understood mechanism. The current study was designed to test the hypothesis that the peroxisome proliferator-activated receptor- coactivator 1α (PGC-1α)/mitochondria axis is involved in ADR-induced podocyte injury. Using MPC5 immortalized mouse podocytes, the ADR dose-dependently induced the downregulation of nephrin and podocin, cell apoptosis, and mitochondrial dysfunction based on the increase in mitochondrial reactive oxygen species (ROS) production, a decrease in mitochondrial DNA (mtDNA) copy number, and the reduction of mitochondrial membrane potential (MMP) and ATP content. Moreover, ADR treatment also remarkably reduced the expression of PGC-1α, an important regulator of mitochondrial biogenesis and function, in podocytes. Strikingly, PGC-1α overexpression markedly attenuated mitochondrial dysfunction, reduction of nephrin and podocin, and the apoptotic response in podocytes following ADR treatment. Moreover, the downregulation of PGC-1α and mitochondria disruption in podocytes were also observed in rat kidneys with ADR administration, suggesting that the PGC-1α/mitochondria axis is relevant to the in vivo ADR-induced podocyte damage. Taken together, these novel findings suggest that dysfunction of the PGC-1α/mitochondria axis is highly involved in ADR-induced podocyte injury. Targeting PGC-1α may be a novel strategy for treating ADR nephropathy and FSGS disease.