<p>Autophagy refers to the intra-cellular metabolism pathways that deliver cytoplasmic target substances to lysosomes for degradation. When cells are exposed to stress; autophagy is developed and has the ability to maintain an adaptable method to the survival of cells. The AMPK- signaling pathway was considered an essential regulator of the autophagy during energy depletion. Many studies suggest that autophagy protects the I/R-induced renal tubular cell injury via an AMPK-regulated pathway. The aim of the current work is to study the effect of metformin on autophagy process and renal alterations associated with ischemia-reperfusion injury through examining its effect on renal functions, oxidative markers and expression of <i>ATG7</i> and <i>LC3II</i> gene in renal tissue. Sham rats (control, <i>n</i> = 10); the rats in the sham group were pre-treated with saline before laparotomy, Group 2 (Ischemic-reperfusion injury, I/R, <i>n</i> = 20); bilateral renal pedicles were clipped for 45 min, followed by perfusion for 24 h to establish I/R model, Group 3(IR + metformin 300 mg/kg, <i>n</i> = 20), Animals were pre-treated with the metformin (Met) at 300 mg/kg 2 doses 2 h, 12 h prior to 45 min of ischemia. The results showed that there is disturbed renal functions as evidenced by the increase in kidney function parameters. Metformin treatment had a protective effect in group that receive treatment through up-regulation of autophagy markers <i>ATG7</i> and <i>LC3II</i> gene &amp; improve kidney function tests, MDA and histological findings. The results suggest that metformin-induced autophagy through activating the AMPK pathway has protection impact against kidney I/R injury.</p>