Cyclosporin A induces apoptosis in rat hepatocytes in culture.
Cyclosporin A (CsA) at concentrations up to 1 microM induced apoptosis in a dose-dependent manner in cultured rat hepatocytes for 48 h in the presence of insulin and epidermal growth factor (EGF). The effect of CsA was evidenced by the DNA fragmentation pattern constituted by fragments of multiples of 180-200 base pairs, which is a characteristic of programmed cell death. The metabolic activity did not change significantly in the presence of 0.1 microM CsA and diminished to 49% of control in the presence of 1 microM CsA. Changes in the metabolic activity were correlated with a decrease in both [methyl-3H]thymidine uptake and DNA content, which reflects a decrease in the cell number. The treatment of cells with CsA (1 microM) decreased the metabolic activity/DNA content ratio by 24% with respect to dimethyl sulphoxide (DMSO) control, which also suggests, under these conditions, that the necrosis achieved is at most only 24%. In addition, the changes observed (apoptotic process, arrest of the cell cycle and apparition of a necrotic process) were correlated with an increase in the high-affinity guanosine triphosphatase (GTPase) enzymes.
Román
I D
ID
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Alcalá de Henares, Spain.
Rodríguez-Henche
N
N
Fueyo
J A
JA
Zueco
J A
JA
Menor
C
C
Prieto
J C
JC
Guijarro
L G
LG
D000165:Acridine Orange / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology*; D004247:DNA / Q000096:biosynthesis; D053938:DNA Fragmentation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004996:Ethidium / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar
D007328:Insulin; D004815:Epidermal Growth Factor; D016572:Cyclosporine; D004247:DNA; D020558:GTP Phosphohydrolases; D004996:Ethidium; D000165:Acridine Orange