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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: Astrocytic gap junction inhibition by carbenoxolone enhances the protective effects of ischemic preconditioning following cerebral ischemia

Fig. 3

Levels of glutamate, reactive oxygen species, and inflammatory factors are affected by oxygen-glucose deprivation. a Cytokine levels. We used an enzyme-linked immunosorbent assay to measure interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and inducible nitric oxide synthase (iNOS) levels, while levels of glutamate were measured using a colorimetric method. These measurements were obtained 6 h following oxygen-glucose deprivation (OGD/R). b Measurement of reactive oxygen species (ROS). Live cell staining for ROS was performed in astrocytes using the ROS sensor reagent CellROX™ Deep Red (red). The astrocyte cell surface marker anti-glutamate-aspartate transporter (GLAST, green) was also used. The images were obtained using deconvolution fluorescence microscopy. All scale bars represent 200 μm. c Measurement of absorbance. We also quantitatively measured ROS expression by collecting absorbance data using a microplate reader at a wave length of 485 nm. IP + 6-diazo-5-oxo-L-norleucine (DON) significantly reduced ROS relative to OGD/R and IP + OGD/R + CBX; IP + OGD/R, compared with OGD/R, exhibited weak statistical for a reduction in ROS. Compared with OGD/R, CBX, or DON reduced the ROS amount of OGD/R significantly. (**P < 0.01; ***P < 0.0001; each scale bar represents 200 μm)

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