Acute kidney injury (AKI) remains a major clinical event with high mortality rates. We previously identified renal miR-182 as the main driver of post-transplantation AKI. Therefore, we tested the causal inference of miR-182 by inhibiting its renal expression in vivo . In 45 rats AKI was induced by right nephrectomy and contralateral clamping of the renal pedicle for 40 minutes. Systemically administered antisense oligonucleotide (ASO) inhibited miR-182 in the kidneys up to 96 hours. The maximum creatinine elevation was on day 2 after injury (ASO 2.5 mg/kg: median, 1.9 mg/dL interquartile range (IQR), 1.3 to 3.2 mg/dL; ASO 25 mg/kg: median, 2.8 mg/dL IQR, 0.7 to 5.0 mg/dL; mismatch oligonucleotide (MM) 25 mg/kg: median, 5.7 mg/dL IQR, 5.0 to 5.8 mg/dL; saline: 4.4 mg/dL IQR, 3.5 to 5.8 mg/dL; P  = 0.016, analysis of variance). Blinded semiquantitative histologic evaluation of renal biopsies showed better preserved morphology in both ASO groups than saline- and MM- treated kidneys in overall injury scores, ASO both concentrations: median, 1 (IQR, 1 to 1); saline: median, 3 (IQR, 3 to 3); MM: median, 3 (IQR, 3 to 3); P  < 0.001, analysis of variance. ASO facilitated cell proliferation, metabolism, and angiogenesis on a genome-wide level. ASO when applied in normothermic kidney machine perfusion reduced renal miR-182 expression by more than two magnitudes. In summary, we showed that in vivo inhibition of miR-182 by ASO improved kidney function and morphology after AKI. This technique may be applicable to reduce the high rate of AKI in the human renal transplantation setting.