Ischemia, and subsequent
acidosis, induces neuronal death following
brain injury. Oxidative stress is believed to be a key component of this neuronal degeneration. Acute chemical
ischemia (
azide in the absence of external
glucose) and
acidosis (external media buffered to pH 6.0) produce increases in intracellular
calcium concentration ([Ca2+]i) and inward membrane currents in cultured rat cortical neurons. Two α-
tocopherol analogues,
trolox and
butylated hydroxytoluene (
BHT), and the spin trapping molecule α-
Phenyl-N-tert-butylnitrone (PBN) were used to determine the role of
free radicals in these responses. PBN and
BHT inhibited the initial transient increases in [Ca2+]i, produced by
ischemia,
acidosis and acidic
ischemia and increased steady state levels in response to
acidosis and the acidic
ischemia.
BHT and PBN also potentiated the rate at which [Ca2+]i increased after the initial transients during acidic
ischemia.
Trolox inhibited peak and sustained increases in [Ca2+]i during
ischemia.
BHT inhibited
ischemia induced initial inward currents and
trolox inhibited initial inward currents activated by
acidosis and acidic
ischemia. Given the inconsistent results obtained using these
antioxidants, it is unlikely their effects were due to elimination of
free radicals. Instead, it appears these compounds have non-specific effects on the
ion channels and exchangers responsible for these responses.