•Tyrosineprotein kinase, GPR15LG, KAZALD1, ADH1B, and thirty metabolites have been identified as potential biomarkers for renal ischemia–reperfusion injury.•Different durations of renal ischemia lead to unique pathological processes and varying components in the venous blood.•During the duration of renal ischemia lasting 40 min, the primary pathological process involves protein digestion and absorption.•During the extended duration of renal ischemia lasting 60 min, the primary pathological mechanisms involve central carbon metabolism, the glucagon signaling pathway, and pyruvate metabolism. Acute kidney injury (AKI) is frequently caused by renal ischemia–reperfusion injury (IRI). Identifying potential renal IRI disease biomarkers would be useful for evaluating AKI severity. We used proteomics and metabolomics to investigate the differences in renal venous blood between ischemic and healthy kidneys in an animal model by identifying differentially expressed proteins (DEPs) and differentially expressed protein metabolites (DEMs). Nine pairs of renal venous blood samples were collected before and at 20, 40, and 60 min post ischemia. The ischemia time of Group A, B and C was 20,40 and 60 min. The proteome and metabolome of renal venous blood were evaluated to establish the differences between renal venous blood before and after ischemia. We identified 79 common DEPs in all samples of Group A, 80 in Group B, and 131 in Group C. Further common DEPs among all three groups were Tyrosineprotein kinase, GPR15LG, KAZALD1, ADH1B. We also identified 81, 64, and 83 common DEMs in each group respectively, in which 30 DEMs were further common to all groups. Bioinformatic analysis of the DEPs and DEMs was conducted. This study demonstrated that different pathological processes occur during short- and long-term renal IRI. Tyrosine protein kinase, GPR15LG, Kazal-type serine peptidase inhibitor domain 1, and all-trans-retinol dehydrogenase are potential biomarkers of renal IRI.