A defect in Nrf2 signaling constitutes a mechanism for cellular stress hypersensitivity in a genetic rat model of type 2 diabetes.

Nrf2 regulates the expression and coordinated induction of a battery of antioxidant phase 2 genes that protect cells against the cumulative damaging effects of oxidative stress (OS), a major contributor in the development of chronic diabetic complications. Using cultured dermal fibroblasts from rats with type 2 diabetes (DFs), we investigated the intracellular redox status and the adaptive response to OS, in which Nrf2 plays a central role. Our data confirmed that the generation of superoxide by NADPH oxidase and the mitochondria was enhanced in DFs compared with corresponding controls. This was associated with a decrease in the antioxidant capacity and an increase in the sensitivity of these DFs to hydrogen peroxide-induced necrotic cell death. Nrf2 levels in total cell extracts were diminished, and this abnormality appears to stem from a diabetes-related decrease in Nrf2 protein stability. Endogenous (oligomycin) and exogenous (tert-butylhydroquinone) induction of OS enhanced the nuclear translocation of Nrf2 and increased the mRNA expression of Nrf2-sensitive genes in control but not DFs. The activity of the GSK-3β/Fyn axis was increased markedly in DFs when compared with the corresponding controls. Chemical inhibition of GSK-3β mitigated the diabetes-related suppression of the OS-induced nuclear accumulation of Nrf2 and the transcriptional activation of the genes downstream of Nrf2. Overall, these findings suggest that an augmentation in GSK-3β/Fyn signaling during diabetes contributes to a deficit in both the cellular redox state and the Nrf2-based adaptive response to OS. Moreover, they may also offer a new perspective in the understanding and treatment of nonhealing diabetic wounds.