The mechanisms of oxidative DNA damage and apoptosis induced by norsalsolinol, an endogenous tetrahydroisoquinoline derivative associated with Parkinson's disease.
Tetrahydroisoquinoline (TIQ) derivatives are putative neurotoxins that may contribute to the degeneration of dopaminergic neurons in Parkinson's disease. One TIQ, norsalsolinol (NorSAL), is present in dopamine-rich areas of human brain, including the substantia nigra. Here, we demonstrate that NorSAL reduces cell viability and induces apoptosis via cytochrome c release and caspase 3 activation in SH-SY5Y human neuroblastoma cells. Cytochrome c release, caspase 3 activation, and apoptosis induction were all inhibited by the antioxidant N-acetylcysteine. Thus, reactive oxygen species (ROS) contribute to apoptosis induced by NorSAL. Treatment with NorSAL also increased levels of oxidative damage to DNA, a stimulus for apoptosis, in SH-SY5Y. To clarify the mechanism of intracellular DNA damage, we examined the DNA damage caused by NorSAL using (32)P-5'-end-labeled isolated DNA fragments. NorSAL induced DNA damage in the presence of Cu(II). Catalase and bathocuproine, a Cu(I) chelator, inhibited this DNA damage, suggesting that ROS such as the Cu(I)-hydroperoxo complex derived from the reaction of H(2)O(2) with Cu(I), promote DNA damage by NorSAL. In summary, NorSAL-generated ROS induced oxidative DNA damage, which led to caspase-dependent apoptosis in neuronal cells.
Kobayashi
Hatasu
H
Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Edobashi 2-174, Tsu, Mie, Japan.
Fukuhara
Kiyoshi
K
Tada-Oikawa
Saeko
S
Yada
Yuki
Y
Hiraku
Yusuke
Y
Murata
Mariko
M
Oikawa
Shinji
S
D000080242:8-Hydroxy-2'-Deoxyguanosine; D017209:Apoptosis / Q000187:drug effects*; D001345:Autoradiography; D016227:Benzoquinones / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D003300:Copper / Q000494:pharmacology; D045304:Cytochromes c / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D009243:NAD / Q000378:metabolism; D009447:Neuroblastoma; D010618:Phenanthrolines / Q000494:pharmacology; D010759:Phosphorus Isotopes / Q000378:metabolism; D012490:Salsoline Alkaloids / Q000494:pharmacology*; D044005:Tetrahydroisoquinolines / Q000737:chemistry; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D016227:Benzoquinones; D016166:Free Radical Scavengers; D010618:Phenanthrolines; D010759:Phosphorus Isotopes; D012490:Salsoline Alkaloids; D044005:Tetrahydroisoquinolines; D013778:Tetrazolium Salts; D013844:Thiazoles; D009243:NAD; C004532:quinone; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; D045304:Cytochromes c; C020784:norsalsolinol; C002478:bathocuproine; D014446:Tyrosine 3-Monooxygenase; D053148:Caspase 3; C022616:thiazolyl blue; D003849:Deoxyguanosine; C029892:cupric chloride