The beneficial effect of purified fraction of sulphated polysaccharide extracted from Sargassum myriocystum (SMP) was examined on the gentamicin-induced nephrotoxicity in adult zebrafish. The major purified fractions (SMP1, SMP2 and SMP3) were obtained by anion-exchange and size-exclusion chromatography and characterized by FTIR, GC[sbnd]MS and NMR. The in vitro antioxidant activities of all purified SMP fractions were analysed. The SMP2 showed maximum carbohydrate, sulphate and fucose content with strong antioxidant activity than other fractions. Further, we evaluated the efficacy of SMP2 against gentamicin-induced nephrotoxicity in zebrafish model. The SMP2 administered group showed a significant attenuation in oxidative stress and histopathological alterations observed in renal tissues of gentamicin treated group. Moreover, the SMP2 supressed renal mRNA expression levels of KIM-1, NF-κB, TNF-α and IL-6 in dose-dependent manner. Thus, the present study suggests that the SMP2 is a potent antioxidant with anti-inflammatory and renoprotective properties that ameliorated the GEN induced nephrotoxicity in adult zebrafish. © 2019 Elsevier B.V.