Figure 3

Cytoprotective potential of Cat-sulf and Pyr-sulf. (a) HBMEC line submitted to oxidative stress (300 µM H₂O₂); (b) primary mouse cerebellar granule cells exposed to glutamate excitotoxicity (100 µM glutamate); (c) 3D aggregates containing neurons and astrocytes exposed to oxidative injury (300 µM t-BHP). Cells were pre-incubated with 5 µM of each bioavailable polyphenol metabolite for 24 h and then injured with the respective lesion. Cell viability was assessed and is presented as percentage relatively to control. Statistical differences are denoted as ***p < 0.001, **p < 0.01 and *p < 0.05 relatively to control and as ^###p < 0.001, ^##p < 0.01 and ^# p< 0.05 relatively to each lesion (H₂O₂, glutamate or t-BHP). (d-f) Alterations in protein markers of the neuronal (β-III tubulin) and astrocytic (GFAP) population of 3D aggregates towards the t-BHP lesion without and with pre-incubation with idebenone (Ide), a control drug, and with Pyr-sulf. (d) Representative western blot and (e) β-III tubulin and (f) GFAP fold changes in protein levels normalized to GAPDH. Statistical differences are denoted as ***p < 0.001, **p < 0.01 and *p < 0.05 relatively to control and as ^###p < 0.001 relatively to t-BHP. Western blots were analyzed under the same experimental conditions. Data are presented as the means ± SD, n = 3.