RTA-408 Protects Kidney from Ischemia-Reperfusion Injury in Mice via Activating Nrf2 and Downstream GSH Biosynthesis Gene

Figure 7

RTA-408 protected HK-2 cells from hypoxia-reoxygenation (H/R) injury, but GCL inhibitor BSO can blunt the protection effect. (a) Dose-dependent protective effect of RTA-408 on H/R-treated HK2 cells. HK2 cells were pretreated with vehicle (DMSO) or RTA-408 (1 nM,10 nM, and 100 nM) for 24 h and were exposed to H/R (24 h hypoxia and 6 h reoxygenation) or not, at which time they were harvested and assessed using a LDH (cell injury marker) assay; each value was expressed as mean ± SD (). ; ^# versus the 0 μM + H/R group. (b) HK2 cells, pretreated with vehicle (DMSO) or RTA-408 (100 nM) for 24 h, were exposed to H/R. The CON group which did not receive any treatment served as the control. Relative mRNA expression of Nrf2 (), HO-1 (), and GCLc () was assessed. Values were expressed as mean ± SD. versus the DMSO + H/R group. (c, d) GCLc immunofluorescence in RTA-408- (100 nM) and H/R-treated cells. The quantitated relative mean density of immunofluorescent was expressed as mean ± SD (). versus the DMSO + H/R group. (e) GCL inhibitor BSO partly abolished RTA-408’s protective effect against H/R. Cell viability was assessed after the different treatment combination of H/R, RTA-408 (100 mM), and L-buthionine sulfoximine (1 mM, BSO, Sigma). Data were expressed as mean ± SD (). versus the DMSO + H/R group. ^# versus the 408 + H/R group.

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