Ischemia/Reperfusion (I/R) injury contributes to acute kidney injury (AKI) and subsequent chronic kidney disease (CKD). I/R injury model by single clamping renal artery is usually used. I confirmed repeated I/R injury after right nephrectomy leads to glomerular injury. In the present study, the aim of study is to produce the new repeated I/R model before right nephrectomy. The rats were divided four groups, sham, single I/R, twice I/R and three time I/R. Animals were anesthetized by medetomidine, midazolam and butorphanol and the left artery and vein were clamped for 45 minutes. A right kidney was removed a week after final I/R. Animals were sacrificed after 8 weeks and we measured renal functional parameters. Rats of all groups were alive. Urinary excretion of protein was progressively increased in all I/R although the other renal functional parameters were not changed. Increment of urinary protein in nephrectomy after repeated I/R seemed to be as large as that in repeated I/R after nephrectomy. In conclusion, repeated I/R-injury model is possibly useful for investigation of AKI to CKD transition.