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b-Amyloid peptide (Ab) is the main component of senile plaques in patients with Alzheimer's disease, and is known to be a main pathogenic factor of the disease. Recent evidence indicates that activation of NADPH oxidase (NOX) in microglia or astrocytes may be a source of Ab-induced reactive oxygen species (ROS). We investigated the role of neuronal NOX in Ab-induced neuronal death in mouse mixed cortical cultures. Cell death was assessed by measuring lactate dehydrogenase efflux to bathing media 24 or 48 hr after exposure to Ab25-35, a fragment of Ab with an equivalent neurotoxic effect. Ab25-35 induced neuronal death in concentration- and time- dependent manners with apoptotic features. Neuronal death was significantly attenuated, not only by anti-apoptotic drugs, such as z-VAD-fmk and cycloheximide, but also by antioxidants, such as trolox, ascorbic acid, and epigallocatethin gallate. We also demonstrated that treatment with 20 mM Ab25-35 increased fluorescent signals in mixed cortical cultures, but produced only weak signals in pure astrocyte cultures in the presence of 2’,7’-dichlorofluorescin diacetate (DCF-DA), an indicator for intracellular ROS. Increased DCF-DA fluorescence was markedly inhibited, not only by trolox, but also by selective NOX inhibitors, such as apocynin and AEBSF. Western blot analyses revealed that Ab25-35 increased the expression of gp91phox, a main subunit of NOX in cells. The above antioxidants, apocynin, and AEBSF significantly attenuated neuronal death induced by Ab25-35. Furthermore, the gp91phox-specific siRNA-based knockdown of NOX significantly inhibited neuronal death. These results suggest that activation of neuronal NOX is involved in Ab25-35-induced neuronal death.
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참고문헌 (30)
참고문헌 신청
Mattson MP / 2000 / Apoptosis in neurodegenerative disorders / Nat Rev Mol Cell Biol 1:120~129
Small DH / 1999 / Alzheimer’s disease and the amyloid beta protein: what is the role of amyloid? / J Neurochem 73:443~449
Small DH / 2001 / Alzheimer's disease and Abeta toxicity : from top to bottom / Nat Rev Neurosci 2:595~598
Verdier Y / 2004 / Amyloid beta-peptide interactions with neuronal and glial cell plasma membrane : binding sites and implications for Alzheimer’s disease / J Pept Sci 10:229~248
Mattson MP / 1997 / Cellular actions of beta-amyloid precursor protein and its soluble and fibrillogenic derivatives / Physiol Rev 77:1081~1132
Small DH / 1999 / Alzheimer’s disease and the amyloid beta protein: what is the role of amyloid? / J Neurochem 73:443~449
Small DH / 2001 / Alzheimer's disease and Abeta toxicity : from top to bottom / Nat Rev Neurosci 2:595~598
Verdier Y / 2004 / Amyloid beta-peptide interactions with neuronal and glial cell plasma membrane : binding sites and implications for Alzheimer’s disease / J Pept Sci 10:229~248
Mattson MP / 1997 / Cellular actions of beta-amyloid precursor protein and its soluble and fibrillogenic derivatives / Physiol Rev 77:1081~1132
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