Acute kidney injury (AKI) is a public health problem associated with significant morbidity and mortality, and is still a major challenge for nephrologists. This work aimed to evaluate the role of autophagy activation in the prevention of cisplatin-induced acute kidney injury. This experimental work included 50 male Sprague–Dawley rats that were 60–80 days old. The rats were divided into five groups randomly. We used rapamycin, a well-known activator of autophagy, and chloroquine, a known agent that inhibits autophagy flux. By quantifying serum creatinine, we assessed the impact of rapamycin on kidney function after exposure to the nephrotoxic agent cisplatin. To assess autophagy induction, we used the LC3-II antibody. As regards the use of rapamycin, rats received cisplatin plus rapamycin at 2 mg/kg daily intraperitoneal. The change in serum creatinine was statistically significantly higher than that in rats that received cisplatin only (P = 0.028). With the use of chloroquine, we found that rats received cisplatin plus rapamycin plus chloroquine, serum creatinine was statistically significantly lower than that in rats that received cisplatin plus rapamycin (P = 0.001) and also lower than that in rats that received cisplatin only (P = 0.015). Also, histopathological data in rats that received cisplatin plus rapamycin plus chloroquine revealed a lower ATN score than cisplatin alone (P = 0.001). We can conclude that use of rapamycin didn’t offer nephroprotection while chloroquine use attenuated the level of acute kidney injury.