Cultured rat vascular smooth muscle cells are resistant to methylamine toxicity: no correlation to semicarbazide-sensitive amine oxidase.
Methylamine (MA), a component of serum and a metabolite of nicotine and certain insecticides and herbicides, is metabolized by semicarbazide-sensitive amine oxidase (SSAO). MA is toxic to cultured human umbilical vein and calf pulmonary artery endothelial cells. Endothelial cells, which do not exhibit endogenous SSAO activity, are exposed to SSAO circulating in serum. In contrast, vascular smooth muscle cells (VSMC) do exhibit innate SSAO activity both in vivo and in vitro. This property, together with the critical localization of VSMC within the arterial wall, led us to investigate the potential toxicity of MA to VSMC. Cultured rat VSMC were treated with MA (10-5 to 1 M). In some cultures, SSAO was selectively inhibited with semicarbazide or MDL-72145 [(E)-2-(3,4-dimethoxyphenyl)-3-fluoroallylamine]. Cytotoxicity was measured via MTT, vital dye exclusion, and clonogenic assays. MA proved to be toxic to VSMC only at relatively high concentrations (LC(50) of 0.1 M). The inhibition of SSAO with semicarbazide or MDL-72145 did not increase MA toxicity, suggesting that the production of formaldehyde via tissue-bound, SSAO-mediated MA metabolism does not play a role in the minimal toxicity observed in isolated rat VSMC. The omission of fetal calf serum (FCS), which contains high SSAO activity, from media similarly showed little effect on cytotoxicity. We conclude that VSMC--in contrast to previous results in endothelial cells--are relatively resistant to MA toxicity, and SSAO does not play a role in VSMC injury by MA.
Langford
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SD
Flight Definition, Wyle Laboratories, NASA Lyndon B. Johnson Space Center, Houston, TX, USA.
Trent
M B
MB
Boor
P J
PJ
D006631:Amine Oxidase (Copper-Containing) / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D002999:Clone Cells; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D005557:Formaldehyde / Q000633:toxicity; D008744:Methylamines / Q000633:toxicity*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013778:Tetrazolium Salts / Q000633:toxicity; D013844:Thiazoles / Q000633:toxicity
D016895:Culture Media, Serum-Free; D008744:Methylamines; D013778:Tetrazolium Salts; D013844:Thiazoles; D005557:Formaldehyde; C027451:methylamine; D006631:Amine Oxidase (Copper-Containing); C022616:thiazolyl blue