alpha-Asarone toxicity in long-term cultures of adult rat hepatocytes.
In this work we studied the toxic effects of alpha-asarone, a hypolipidemic active principle of Guatteria gaumeri Greenman, on long-term cultures of adult rat hepatocytes cultivated on a feeder layer of 3T3 cells. The exposure for one and two weeks to alpha-asarone (1-50 micrograms/ml) produced intracytoplasmic lipid droplets and at higher concentrations (25-50 micrograms/ml) retraction of the hepatocyte cords and cell detachment. Ultrastructurally, the treated cultures (10 micrograms/ml) showed enlargement and vacuolization of the mitochondria in addition to lipid droplets. The triacylglycerol content increased up to 2.3-fold in the cultures treated for one week with 50 micrograms/ml, whereas the protein content per culture, a rough estimate of cell number and viability, decreased by up to 53% in the cultures treated for two weeks with 50 micrograms/ml. The synthesis and secretion of proteins, measured by the incorporation of [3H]-leucine into cellular and secreted macromolecules, decreased also in the cultures exposed. After one and two week exposure to 50 micrograms/ml of alpha-asarone, the secretion of labeled proteins decreased by 53 and 67%, respectively, whereas the synthesis of cellular and total proteins decreased by 48-67%, respectively. The secretion of proteins was the most sensitive parameter of alpha-asarone toxicity. The mean inhibitory dose (ID50), i.e, that producing 50% inhibition in the incorporation of the labeled precursor, was 22.12 and 5.04 micrograms/ml after one and two weeks exposure, respectively. Our results show that long-term exposure to micromolar concentrations of alpha-asarone produces morphologic and ultrastructural alterations, triacylglycerol accumulation (fatty liver), and inhibition of protein synthesis and secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
López
M L
ML
Pharmacology and Toxicology Department, Centro de Investigación y Estudios Avanzados del IPN, México, D.F.
Hernández
A
A
Chamorro
G
G
Mendoza-Figueroa
T
T
D016475:3T3 Cells; D000081248:Allylbenzene Derivatives; D000818:Animals; D000840:Anisoles / Q000633:toxicity*; D002273:Carcinogens / Q000633:toxicity; D002478:Cells, Cultured; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D008297:Male; D051379:Mice; D010946:Plants, Medicinal / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar
D000081248:Allylbenzene Derivatives; D000840:Anisoles; D002273:Carcinogens; C012195:asarone