An evaluation of the effects of photoactivation of bithionol, amiodarone and chlorpromazine on human keratinocytes in vitro.
Human skin is a continual target for chemical toxicity, due to its constant exposure to xenobiotics. The skin possesses a number of protective antioxidant systems, including glutathione and enzymic pathways, which are capable of neutralising reactive oxygen species (ROS). In combination with certain chemicals, the presence of ROS might augment the levels of toxicity, due to photoactivation of the chemical or, alternatively, due to an oxidatively-stressed state in the skin which existed prior to exposure to the chemical. Bithionol is a phototoxic anti-parasitic compound. The mechanism of its toxicity and the possible methods of protection from its damaging effects have been explored. The capacity of keratinocytes to protect themselves from bithionol and other phototoxic chemicals has been investigated. In addition, the potential of endogenous antioxidants, such as vitamin C and E, to afford protection to the cells, has been evaluated. The intracellular glutathione stores of HaCaT keratinocytes were reduced following treatment with biothionol. Following photoactivation, both bithionol and chlorpromazine had similar effects, which suggests that glutathione is important in the detoxification pathway of these chemicals. This was confirmed by means of the visual identification of fluorescently-labelled glutathione. Endogenous antioxidants were unable to protect the HaCaT keratinocytes from bithionol toxicity or chlorpromazine phototoxicity. Amiodarone was shown to have no effect on cellular glutathione levels, which suggests that an alternative mechanism of detoxification was occurring in this case. This was supported by evidence of the protection of HaCaT cells from amiodarone phototoxicity via endogenous antioxidants. Thus, it appears that amiodarone toxicity is dependent on the levels of non-gluathione antioxidants present, whilst bithionol and chlorpromazine detoxification relies on the glutathione antioxidant system. This type of approach could indicate the likely mechanisms of phototoxicity of chemicals in vitro, with relevance to potential effects in vivo.
Reid
Linzi
L
FRAME Alternatives Laboratory, School of Biomedical Sciences, University of Nottingham, Nottingham, UK.
Khammo
Nancy
N
Clothier
Richard H
RH
D000638:Amiodarone / Q000633:toxicity*; D000826:Animal Testing Alternatives; D000889:Anti-Arrhythmia Agents / Q000633:toxicity*; D000891:Anti-Infective Agents, Local / Q000633:toxicity*; D000975:Antioxidants / Q000494:pharmacology; D001735:Bithionol / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002746:Chlorpromazine / Q000633:toxicity; D017484:Dermatitis, Phototoxic / Q000209:etiology; D018492:Dopamine Antagonists / Q000633:toxicity; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects; D008564:Membrane Potentials / Q000187:drug effects / Q000528:radiation effects; D008928:Mitochondria / Q000187:drug effects / Q000528:radiation effects; D010084:Oxidation-Reduction / Q000187:drug effects / Q000528:radiation effects; D018384:Oxidative Stress / Q000187:drug effects / Q000528:radiation effects; D014466:Ultraviolet Rays
D000889:Anti-Arrhythmia Agents; D000891:Anti-Infective Agents, Local; D000975:Antioxidants; D018492:Dopamine Antagonists; D005452:Fluoresceins; D005456:Fluorescent Dyes; C069306:5-chloromethylfluorescein; D001735:Bithionol; D005978:Glutathione; D000638:Amiodarone; D002746:Chlorpromazine