Studies on the neurotoxicity of 6,7-dihydroxy-1-methyl-1,2,3,4-tetrahydroisoquinoline (salsolinol) in SH-SY5Y cells.
We have studied the hypothesis that 6,7-dihydroxy-1-methyl-1,2,3,4-tetrahydroisoquinoline (salsolinol) is neurotoxic. Salsolinol induced a significant time and dose related inhibition of 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; thiazoyl blue (MTT) reduction, and increased lactate dehydrogenase release (LDH) release from human SH-SY5Y neuroblastoma cells, at concentrations within the range of 1-methyl-4-phenylpyridinium (MPP+) cytotoxicity, in vitro. Cytotoxicity was not inhibited by the addition of antioxidants, monoamine oxidase inhibitors or imipramine. In confluent monolayers, salsolinol stimulated catecholamine uptake with EC50 values of 17 muM and 11 muM, for noradrenaline and dopamine, respectively. Conversely, at concentrations above 100 muM, salsolinol inhibited the uptake of noradrenaline and dopamine, with IC50 values of 411 muM and 379 muM, respectively. The inhibition of catecholamine uptake corresponded to the increase displacement of [3H]nisoxetine from the uptake 1 site by salsolinol, as the Ki (353 muM) for displacement was similar to the IC50 (411 and 379 muM) for uptake. Salsolinol stimulated catecholamine uptake does not involve the uptake recognition site, or elevation of cAMP, cGMP, or inhibition of protein kinase C. Salsolinol also inhibited both carbachol (1 mM) and K+ (100 mM, Na+ adjusted) evoked released of noradrenaline from SH-SY5Y cells, with IC50 values of 500 muM and 120 muM, respectively. In conclusion, salsolinol appears to be cytotoxic to SH-SY5Y cells, via a mechanism that does not require uptake 1, bioactivation by monoamine oxidase, or membrane based free radical damage. The effects of salsolinol on catecholamine uptake, and the mechanism of toxicity require further investigation.
Willets
J M
JM
Division of Chemical Pathology, C.M.H.T., University of Leicester, UK.
Lambert
D G
DG
Lunec
J
J
Griffiths
H R
HR
D000262:Adenylyl Cyclases / Q000819:agonists; D018759:Adrenergic Uptake Inhibitors / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism*; D016923:Cell Death; D003010:Clorgyline / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006162:Guanylate Cyclase / Q000819:agonists; D006801:Humans; D007099:Imipramine / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007546:Isoquinolines / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008996:Monoamine Oxidase Inhibitors / Q000494:pharmacology; D009447:Neuroblastoma; D009498:Neurotoxins / Q000633:toxicity*; D009599:Nitroprusside / Q000494:pharmacology; D011976:Receptors, Muscarinic / Q000378:metabolism; D012642:Selegiline / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D018759:Adrenergic Uptake Inhibitors; D002395:Catecholamines; D007211:Indoles; D007546:Isoquinolines; D008996:Monoamine Oxidase Inhibitors; D009498:Neurotoxins; D011976:Receptors, Muscarinic; D009599:Nitroprusside; D012642:Selegiline; C036617:salsolinol; D007770:L-Lactate Dehydrogenase; D000262:Adenylyl Cyclases; D006162:Guanylate Cyclase; D003010:Clorgyline; D007099:Imipramine; C064758:Ro 31-8220