Retinol, at concentrations greater than the physiological limit, induces oxidative stress and apoptosis in human dermal fibroblasts.
We have investigated the dose (in the range of microM) and time-dependent effects of four different retinoids (retinol, retinal, retinoic acid and retinol palmitate) on human dermal fibroblasts cultivated in vitro. Retinol and retinal, at a concentration of 20 microM, caused cell damage as evaluated by lactate dehydrogenase activity released into the culture medium. The oxidised glutathione (GSSG)/reduced glutathione (GSH) ratio and malondialdehyde production indicated that 20 microM of retinol provoked oxidative stress in the cultivated human fibroblasts. In the first 8 h after retinol treatment the levels of p53 and Bax proteins as well as caspase 3 activity increased, suggesting apoptotic cell death during the first hours of treatment. If the retinol treatment exceeded 18-24 h we observed necrotic cell death. Vitamin E and coenzyme Q(10) had a protective effect against oxidative stress generated by retinol. Both antioxidant molecules reduced retinol uptake, and in the case of vitamin E the expression of CRABP-II mRNA was induced, providing a plausible explanation for its protective effect.
Departamento de Bioquímica y Biología Molecular, Facultades de Farmacia y Medicina, Universidad de Valencia, Aveñida Vicent Andrés Estellés s/n, E-46100-Burjassot, Valencia, Spain.
Amparo
A
Gimeno
Rosa
R
Zaragozá
Inma
I
Vivó-Sesé
Juan R
JR
Viña
Vicente J
VJ
Miralles
D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D005978:Glutathione / Q000378:metabolism; D019803:Glutathione Disulfide / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects*; D012176:Retinoids / Q000494:pharmacology*; D012867:Skin / Q000187:drug effects*; D012879:Skin Physiological Phenomena; D014212:Tretinoin / Q000494:pharmacology; D014801:Vitamin A / Q000494:pharmacology*
D000975:Antioxidants; D012176:Retinoids; D014801:Vitamin A; D014212:Tretinoin; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D019803:Glutathione Disulfide