Photodynamic effects of protoporphyrin on the cellular level-an in vitro approach.
The purpose of this study was characterizing the phototoxic action of protoporphyrin and cellular protection mechanisms, as studied on the cellular level. In this process, active oxygen is involved. As a biological system, rat hepatocyte short-term and primary cultures were used. Phototoxicity of protoporphyrin could be observed, after previous absorption of protoporphyrin to membrane structures. Damaging of several cell organelles occurred, such as mitochondria and lysosomes. Peroxisomes were not affected. Coated vesicles located at the periphery of the cells' interior suggested that protoporphyrin absorption is mediated by an active uptake (endocytosis), as well as passive diffusion. Lipid peroxidation played a role in protoporphyrin phototoxicity. Cellular protection mechanisms such as superoxide dismutase and the scavenger glutathione (GSH) protected the cells from active oxygen toxicity. In conclusion, protoporphyrin entered the cells by diffusion and endocytosis. Previous adsorption to the membrane structures was necessary for the expression of protoporphyrin phototoxicity. However, active oxygen itself could not be demonstrated. Lipid peroxidation was involved in cell-damaging processes. Mechanisms of protoporphyrin phototoxicity on the cellular level were studied. Rat hepatocyte primary and short-term cultures proved to be suitable in vitro systems for studying biochemical and morphological effects on the cellular level.
Van Graft
M
M
Department of Veterinary Pharmacology, Pharmacy and Toxicology, Utrecht University, The Netherlands.
Boot
J H
JH
D000818:Animals; D002478:Cells, Cultured; D004050:Ditiocarb / Q000494:pharmacology; D008027:Light; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D008297:Male; D017319:Photosensitizing Agents / Q000633:toxicity*; D011524:Protoporphyrins / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D013482:Superoxide Dismutase / Q000378:metabolism
D017319:Photosensitizing Agents; D011524:Protoporphyrins; D004050:Ditiocarb; D013482:Superoxide Dismutase