The hepatotoxicity of rhein involves impairment of mitochondrial functions.
Metabolism of rhein (4,5-dihydroxyanthraquinone-2-carboxylic acid) in primary cultures of rat hepatocytes caused production of oxygen-derived free radicals by redox cycling; this was shown as an increased rate of superoxide-dismutasesensitive NAD(P)H oxidation and NAD(P)H-cytochrome c reduction. Furthermore, rhein caused a depletion of intracellular reduced glutathione and an immediate, almost 10-fold increase in intracellular free Ca2+. Exposure to rhein also induced the following: a decrease in the mitochondrial membrane potential, as analyzed by uptake of rhodamine 123 (Rh 123); initiation of lipid peroxidation, measured as accumulation of malondialdehyde and 4-hydroxyalkenals; and cell death (LD50 = 20 microM). Pretreatment of cell cultures with dithiothreitol (DTT), nifedipin or N',N'-diphenyl-p-phenylenediamine (DPPD) increased the intracellular free Ca2+ concentration 5-fold but inhibited rhein-induced cytotoxicity. Moreover, addition of these protecting substances maintained the level of ATP and glutathione (GSH) and prevented accumulation of lipid peroxidation products. Depletion of intracellular glutathione by pretreatment with buthionine sulfoximine (BSO), or inhibition of glutathione reductase with 1,3-bis-2-chloroethyl-1-nitrosourea (BCNU) decreased cell viability (LD50 = 2.5 microM). On the other hand, increasing GSH by pretreatment with L-2-oxothiazolidine-4-carboxylic acid (OTC) did not provide complete protection. In summary, rhein undergoes redox cycling that gives rise to oxygen metabolites that affect the mitochondrial membranes (recorded as a decreased membrane potential) and after the plasma membrane (i.e. induced the formation of surface blebs). Mitochondrial malfunction also causes changes in Ca2+ homeostasis and depletion of ATP, which eventually lead to cell death.
Bironaite
D
D
Institute of Biochemistry, Lithuanian Academy of Sciences, Vilnius, Lithuania.
Ollinger
K
K
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D000880:Anthraquinones / Q000633:toxicity*; D000975:Antioxidants / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000633:toxicity*; D005978:Glutathione / Q000032:analysis; D006706:Homeostasis / Q000502:physiology; D015227:Lipid Peroxidation; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D016877:Oxidants / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D000880:Anthraquinones; D000975:Antioxidants; D004791:Enzyme Inhibitors; D016877:Oxidants; D000255:Adenosine Triphosphate; D005978:Glutathione; D002118:Calcium; C020491:rhein