Research Article

Nitro-Oxidative Stress after Neuronal Ischemia Induces Protein Nitrotyrosination and Cell Death

Figure 1

In vitro ischemia induces nitro-oxidative stress. (a) A representative image of nitrotyrosination (bottom image, stained in red) and bright field (top image) in a section of cortex from a stroke brain analyzed by immunohistochemistry. The arrows indicate blood vessels. (b) Primary hippocampal cells were subjected to in vitro ischemia followed by reperfusion with normal medium for 24 h. Nitrotyrosine (red staining) was demonstrated by immunostaining. Nuclei are stained in blue with To-pro 3. (c) Human neuroblastoma cells were subjected to in vitro ischemia reperfused with normal medium for 24 h. Free radical production was detected by DCF fluorescence and quantified. The mean fluorescence of DCF represents the levels of ROS. Data are mean ± SEM values of 4 independent experiments. versus control. (d) NO production (expressed in picomoles) was measured in neurons, microglia, endothelial, and vascular smooth muscle cells challenged with in vitro ischemia and later reoxygenated with normal growing medium containing glucose at 0, 12, and 24 h. Data are mean ± SEM values of 4 independent experiments. ; versus controls at 0 h.
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