BACKGROUND AND PURPOSE: Apoptotic-like pathways may contribute to brain cell death after @entity1779 . In this study, we used a simplified in vitro model of hemoglobin @entity2359 to map the caspase cascades involved and to document the role of oxidative stress. METHODS: Primary neuronal cultures were obtained from @entity35 cerebral cortex and exposed to hemoglobin to induce cell death. @entity137 was assessed via measurements of mitochondrial viability ( @entity98 assay) and @entity880 dehydrogenase (LDH assay). Activation of @entity51682 was measured by Western blot and enzyme activity assays. Various caspase inhibitors (zVADfmk, zDEVDfmk, zIETDfmk, and zLEHDfmk) were tested for neuroprotective efficacy. The role of oxidative stress was assessed with the use of @entity42015 as a potent scavenger of free radicals. RESULTS: Exposure of primary cortical neurons to hemoglobin induced a dose- and time-dependent @entity137 . Western blots showed upregulation of cleaved caspase-3. Enzyme assays showed an increase in caspase-9-like and caspase-3-like activity. However, caspase inhibition did not result in neuroprotection. In contrast, the free radical scavenger @entity42015 significantly reduced hemoglobin-induced @entity528 . Combination treatment with both @entity42015 and the broad spectrum caspase inhibitor zVADfmk did not yield additional protection. CONCLUSIONS: Upstream and downstream caspases were upregulated after hemoglobin-induced @entity2359 in vitro, but only an antioxidant approach with a potent free radical scavenger significantly improved neuronal survival. These data suggest that in addition to the activation of caspase cascades, parallel pathways of oxidative stress may predominate in this model of hemoglobin @entity2359 .

Hemoglobin-induced XXXX in @entity35 cerebral cortical neurons: caspase activation and oxidative stress.

@entity137 :: ('MESH:D064420', 'Disease') :: ['Cytotoxicity', 'cytotoxicity'] @entity51682 :: ('25402;64044;58918', 'Gene') :: ['caspase-3, -8, and -9'] @entity42015 :: ('MESH:C066382', 'Chemical') :: ['U83836E'] @entity98 :: ('CHEBI:53233', 'Chemical') :: ['MTT'] @entity880 :: ('MESH:D019344', 'Chemical') :: ['lactate'] @entity35 :: ('10116', 'Species') :: ['rat'] @entity528 :: ('MESH:D009410', 'Disease') :: ['neuronal death'] @entity1779 :: ('MESH:D002543', 'Disease') :: ['intracerebral hemorrhage'] @entity2359 :: ('MESH:D020258', 'Disease') :: ['neurotoxicity']
@entity137 :: (MESH:D064420,Disease) :: ['cytotoxicity']
