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Background and Aims Drug-induced acute kidney injury (AKI) is a health problem that increases morbidity and mortality rates as well as costs to health systems. Amphotericin B (AnB) is a widely used antifungal, which its therapy is closely associated with nephrotoxicity. The mechanisms of nephrotoxicity are not yet fully understood and there is a need to search for substances that prevent and / or reverse the associated renal damage. (-) - α-Bisabolol (BIS) has been demonstrated nephroprotective effect due to its antioxidant action. Thus, the aim of this study was to evaluated the effect of BIS as a protective substance on renal cell injury induced by amphotericin B. Method LLC-MK2 tubular renal cells were treated with different concentrations of BIS and AnB and its viability was measured by MTT assay to define the working concentrations of BIS and AnB. Then, cells were pretreated or post treated with BIS (31.25 - 125 μM) and AnB (13.97 μM) for 24 hours to evaluate the protective effect of BIS. Control (CT) was treated with sterile PBS. The experimental groups were further submitted to flow cytometry assays for membrane integrity evaluation using 7-AAD and measurement of cytoplasmic ROS production using DCFH-DA. The experiments were performed in triplicate (n = 3), and the data analyzed by One-Way ANOVA with Bonferroni post-test. Results BIS concentrations from 31.25 to 125 μM were selected, as well as the IC50 for AnB (13.97 ± 1.45 μM) for the furthers assays. Pretreatment with BIS increased cell viability by about 10% (31.25 – 63.31 ± 2.04%; 62.5 – 64.75 ± 2.10%; 125 – 63.92 ± 2.72%) compared to the AnB-only group (49.79 ± 1.25%), which had its cell viability reduced by about 50% (CT – 100.70 ± 0.83%). BIS was also able to increase the whole cell population at concentrations of 62.5 (83.10 ± 1.10%) and 125 μM (84.67 ± 0.98%) when compared to the AnB group (76.54 ± 1.18%). In DCF labeling, it was observed that AnB increased the production of ROS in 20% (IFR = 1.21 ± 0.02) when compared to the control (IFR = 1.00 ± 0.01), and BIS (31.25 μM) was able to reduce in 41% (IFR = 0.79 ± 0.01) when compared to the AnB group. Conclusion The (-)-α-bisabolol was able to prevent the renal cytotoxicity induced by amphotericin.