Protective effect of diosmetin on in vitro cell membrane damage and oxidative stress in cultured rat hepatocytes.
Primary cultures of rat hepatocytes were used to study the effects of the flavonoids diosmin and its main metabolite diosmetin on the cell membrane damage caused by erythromycin estolate (EE) and oxidative stress caused by tert-butylhydroperoxide (TBHP). The damage was evaluated by the leakage of intracellular enzymes lactate dehydrogenase, aspartate-aminotransferase and the residual cell content of a lysosomal marker acid phosphatase (AP). After treating the cells for 40 h with diosmetin EE induced less enzyme leakage. The content of AP was kept higher by diosmetin pretreatment after 6 h exposure to EE. Diosmin at the same concentrations had barely any effect. Diosmetin, but not diosmin, also protected against TBHP toxicity and this was related to lower lipid peroxidation and higher glutathione content caused by pretreatment with the flavonoid. When the cells were treated simultaneously with TBHP and diosmetin after 21 h of culture, the protection by the flavonoid was even higher. In fact the antioxidant activity of diosmetin was considerably greater than that of diosmin. After 40 h exposure to both flavonoids diosmin but not diosmetin was detectable in the cell membrane fraction, suggesting that the latter's protective effect is associated with its metabolites.
Villa
P
P
C.N.R. (National Research Council) Center of Cytopharmacology, Department of Pharmacology, Milan, Italy.
Cova
D
D
De Francesco
L
L
Guaitani
A
A
Palladini
G
G
Perego
R
R
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D001219:Aspartate Aminotransferases / Q000032:analysis; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D004145:Diosmin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004918:Erythromycin Estolate / Q000633:toxicity*; D005419:Flavonoids / Q000494:pharmacology*; D005978:Glutathione / Q000032:analysis; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D016877:Oxidants / Q000633:toxicity; D010545:Peroxides / Q000633:toxicity*; D051381:Rats; D020122:tert-Butylhydroperoxide
D005419:Flavonoids; D016877:Oxidants; D010545:Peroxides; D004145:Diosmin; D020122:tert-Butylhydroperoxide; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000135:Acid Phosphatase; D005978:Glutathione; C039602:diosmetin; D004918:Erythromycin Estolate