Irniine, a pyrrolidine alkaloid, isolated from Arisarum vulgare can induce apoptosis and/or necrosis in rat hepatocyte cultures.
The effects of irniine, a pyrrolidine alkaloid extracted from the tubers of Arisarum vulgare, on rat hepatocyte primary cultures and rat liver epithelial cell line (RLEC) were studied. Cytotoxicity was first evaluated by LDH release, MTT and NR tests and MDA production, while cellular alterations were visualized by electron microscopy and DNA gel-electrophoresis. In hepatocyte and RLEC cultures, a major toxicity appeared at 40 microM of irniine and was demonstrated by an increase in LDH release and decreases in MTT reduction and NR uptake while concentrations lower than 40 microM did not induce significant changes in these parameters. However, we observed an increase in MDA production at 30 microM. Important alterations of the nuclei and mitochondria were also visualized by electron microscopy in cells treated with 50 microM. Using DNA gel-electrophoresis, we demonstrated that irniine at 40 and 50 microM induced DNA damage. All together these results demonstrate that: (1) Irniine induces a significant hepatotoxicity. (2) Irniine toxicity is not mediated by a metabolic derivative since RLEC, which do not contain a monooxygenase system, were also affected by this compound. (3) Irniine induces a significant DNA damage and oxidative stress which leads to cell death by necrosis and/or by apoptosis. Moreover, our data suggest that the alkaloid irniine contained in A. vulgare may be involved in the toxic symptoms observed after medicinal use or consumption of the plant tubers as food both by humans and animals.
Rakba
N
N
INSERM U522, Régulations des Equilibres Fonctionnels du Foie Normal et Pathologique, Hôpital Pontchaillou, Rennes, France. nafissa.rakba@rennes.inserm.fr
Melhaoui
A
A
Rissel
M
M
Morel
I
I
Loyer
P
P
Lescoat
G
G
D000470:Alkaloids / Q000302:isolation & purification / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis / Q000187:drug effects; D004249:DNA Damage; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000648:ultrastructure; D009336:Necrosis; D009499:Neutral Red / Q000378:metabolism; D010947:Plants, Toxic / Q000737:chemistry / Q000633:toxicity*; D011759:Pyrrolidines / Q000302:isolation & purification / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D000470:Alkaloids; D011759:Pyrrolidines; D013778:Tetrazolium Salts; D013844:Thiazoles; C411262:irniine; D009499:Neutral Red; D008315:Malondialdehyde; D004247:DNA; D007770:L-Lactate Dehydrogenase; C022616:thiazolyl blue