RESUMO
All-trans-retinal (atRAL) is a highly reactive carbonyl specie, known for its reactivity on cellular phosphatidylethanolamine in photoreceptor. It is generated by photoisomerization of 11-cis-retinal chromophore linked to opsin by the Schiff's base reaction. In ABCA4-associated autosomal recessive Stargardt macular dystrophy, atRAL results in carbonyl and oxidative stress, which leads to bisretinoid A2E, accumulation in the retinal pigment epithelium (RPE). This A2E-accumulation presents as lipofuscin fluorescent pigment, and its photooxidation causes subsequent damage. Here we describe protection against a lethal dose of atRAL in both photoreceptors and RPE in primary cultures by a lipidic polyphenol derivative, an isopropyl-phloroglucinol linked to DHA, referred to as IP-DHA. Next, we addressed the cellular and molecular defence mechanisms in commonly used human ARPE-19 cells. We determined that both polyunsaturated fatty acid and isopropyl substituents bond to phloroglucinol are essential to confer the highest protection. IP-DHA responds rapidly against the toxicity of atRAL and its protective effect persists. This healthy effect of IP-DHA applies to the mitochondrial respiration. IP-DHA also rescues RPE cells subjected to the toxic effects of A2E after blue light exposure. Together, our findings suggest that the beneficial role of IP-DHA in retinal cells involves both anti-carbonyl and anti-oxidative capacities.
Assuntos
Desidroepiandrosterona/farmacologia , Floroglucinol/farmacologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Retinaldeído/toxicidade , Animais , Antioxidantes/farmacologia , Catalase/metabolismo , Linhagem Celular , Sobrevivência Celular , Humanos , Lipofuscina/química , Camundongos , Mitocôndrias/metabolismo , Neurônios/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Oxigênio/química , Consumo de Oxigênio , Fenol/química , Floroglucinol/química , Pigmentação , Substâncias Protetoras/farmacologia , Ratos , Espécies Reativas de Oxigênio , Epitélio Pigmentado da Retina/metabolismo , Retinoides/metabolismo , Relação Estrutura-AtividadeRESUMO
The chemical- and photo- toxicity of chromophore retinal on cells have long been debated. Although we recently showed that retinal and blue light exposure interrupt cellular signaling, a comprehensive study examining molecular underpinnings of this perturbation and its consequences to cellular fate is lacking. Here, we report molecular evidence for blue light excited-retinal induced oxidative damage of polyunsaturated lipid anchors in membrane-interacting signaling molecules and DNA damage in cells using live-cell imaging and in vitro experimentation. The incurred molecular damage irreversibly disrupted subcellular localization of these molecules, a crucial criterion for their signaling. We further show retinal accumulation in lipid-bilayers of cell membranes could enhance the lifetime of retinal in cells. Comparative response-signatures suggest that retinal triggers reactions upon photoexcitation similar to photodynamic therapy agents and generate reactive oxygen species in cells. Additionally, data also shows that exposing retinal-containing cells to sunlight induces substantial cytotoxicity. Collectively, our results explain a likely in vivo mechanism and reaction conditions under which bio-available retinal in physiological light conditions damages cells.
Assuntos
Luz/efeitos adversos , Retinaldeído/toxicidade , Dano ao DNA , Células HeLa , Humanos , Peroxidação de Lipídeos , Estresse Oxidativo , Processos Fotoquímicos , Espécies Reativas de Oxigênio/metabolismoRESUMO
The retinoid cycle is a metabolic process in the vertebrate retina that continuously regenerates 11-cis-retinal (11-cisRAL) from the all-trans-retinal (atRAL) isomer. atRAL accumulation can cause photoreceptor degeneration and irreversible visual dysfunction associated with incurable blinding retinal diseases, such as Stargardt disease, retinitis pigmentosa (RP), and atrophic age-related macular degeneration (AMD). The underlying cellular mechanisms leading to retinal degeneration remain uncertain, although previous studies have shown that atRAL promotes calcium influx associated with cell apoptosis. To identify compounds that mitigate the effects of atRAL toxicity, here we developed an unbiased and robust image-based assay that can detect changes in intracellular calcium levels in U2OS cells. Using our assay in a high-throughput screen of 2,400 compounds, we noted that selective estrogen receptor modulators (SERMs) potently stabilize intracellular calcium and thereby counteract atRAL-induced toxicity. In a light-induced retinal degeneration mouse model (Abca4-/-Rdh8-/-), raloxifene (a benzothiophene-type scaffold SERM) prevented the onset of photoreceptor apoptosis and thus protected the retina from degeneration. The minor structural differences between raloxifene and one of its derivatives (Y 134) had a major impact on calcium homeostasis after atRAL exposure in vitro, and we verified this differential impact in vivo In summary, the SERM raloxifene has structural and functional neuroprotective effects in the retina. We propose that the highly sensitive image-based assay developed here could be applied for the discovery of additional drug candidates preventing photoreceptor degeneration.
Assuntos
Células Fotorreceptoras de Vertebrados/citologia , Substâncias Protetoras/farmacologia , Cloridrato de Raloxifeno/farmacologia , Degeneração Retiniana/prevenção & controle , Epitélio Pigmentado da Retina/citologia , Retinaldeído/toxicidade , Moduladores Seletivos de Receptor Estrogênico/farmacologia , Transportadores de Cassetes de Ligação de ATP/fisiologia , Oxirredutases do Álcool/fisiologia , Animais , Feminino , Masculino , Camundongos , Camundongos Knockout , Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Células Fotorreceptoras de Vertebrados/metabolismo , Degeneração Retiniana/induzido quimicamente , Degeneração Retiniana/patologia , Epitélio Pigmentado da Retina/efeitos dos fármacosRESUMO
It is estimated that abnormal accumulation of all-trans-retinal (atRAL) is a leading cause of photoreceptor degeneration in retinal degenerative diseases. Deficiency of interphotoreceptor retinoid-binding protein (IRBP), a retinoid transporter in the visual cycle, is responsible for the impaired clearance of atRAL and results in atRAL toxicity in retina. Therefore, IRBP has been proposed to be a potent target in preventing atRAL-induced photoreceptor degeneration. In this study, the neuroprotective effect of tetramethylpyrazine (TMP) against atRAL toxicity in the differentiated Y-79 cells, a in vitro model of photoreceptor, was first investigated. Our findings showed that atRAL could induce cytotoxicity, oxidative/nitrosative stresses, apoptosis and leukostasis in the differentiated Y-79 cells; however, the pre-treatment of TMP significantly attenuated such effects in a dose-dependent manner. Furthermore, our results indicated that TMP exerted its neuroprotective effect mainly through upregulating IRBP expression. The present study significantly contributes to better understanding the important role of IRBP in retinal degenerative diseases and forms the basis of the therapeutic development of TMP in such diseases in the future.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Proteínas do Olho/metabolismo , Fármacos Neuroprotetores/farmacologia , Pirazinas/farmacologia , Retinaldeído/toxicidade , Proteínas de Ligação ao Retinol/metabolismo , Regulação para Cima/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Leucostasia/patologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Neuroproteção/efeitos dos fármacos , Nitrosação , Estresse Oxidativo/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
Effective clearance of all-trans-retinal (atRAL) from retinal pigment epithelial (RPE) cells is important for avoiding its cytotoxicity. However, the metabolism of atRAL in RPE cells is poorly clarified. The present study was designed to analyze metabolic products of atRAL and to compare the cytotoxicity of atRAL versus its derivative all-trans-retinal dimer (atRAL-dimer) in human RPE cells. We found that all-trans-retinol (atROL) and a mixture of atRAL condensation metabolites including atRAL-dimer and A2E were generated after incubating RPE cells with atRAL for 6h, and the amount of atRAL-dimer was significantly higher than that of A2E. In the eyes of Rdh8-/- Abca4-/- mice, a mouse model with defects in retinoid cycle that displays some symbolic characteristics of age-related macular degeneration (AMD), the level of atRAL-dimer was increased compared to wild-type mice, and was even much greater than that of A2E & isomers. The cytotoxicity of atRAL-dimer was reduced compared with its precursor atRAL. The latter could provoke intracellular reactive oxygen species (ROS) overproduction, increase the mRNA expression of several oxidative stress related genes (Nrf2, HO-1, and γ-GCSh), and induce ΔΨm loss in RPE cells. By contrast, the abilities of atRAL-dimer to induce intracellular ROS and oxidative stress were much weaker versus that of concentration-matched atRAL, and atRAL-dimer exhibited no toxic effect on mitochondrial function at higher concentrations. In conclusion, the formation of atRAL-dimer during atRAL metabolic process ameliorates the cytotoxicity of atRAL by reducing oxidative stress.
Assuntos
Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Epitélio Pigmentado da Retina/efeitos dos fármacos , Retinaldeído/análogos & derivados , Transportadores de Cassetes de Ligação de ATP/biossíntese , Transportadores de Cassetes de Ligação de ATP/genética , Oxirredutases do Álcool/biossíntese , Oxirredutases do Álcool/genética , Animais , Linhagem Celular , DNA Complementar/biossíntese , DNA Complementar/genética , Humanos , Degeneração Macular/genética , Degeneração Macular/prevenção & controle , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Knockout , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo , Epitélio Pigmentado da Retina/citologia , Retinaldeído/química , Retinaldeído/toxicidadeRESUMO
Among retinal macular diseases, the juvenile recessive Stargardt disease and the age-related degenerative disease arise from carbonyl and oxidative stresses (COS). Both stresses originate from an accumulation of all-trans-retinal (atRAL) and are involved in bisretinoid formation by condensation of atRAL with phosphatidylethanolamine (carbonyl stress) in the photoreceptor and its transformation into lipofuscin bisretinoids (oxidative stress) in the retinal pigment epithelium (RPE). As atRAL and bisretinoid accumulation contribute to RPE and photoreceptor cell death, our goal is to select powerful chemical inhibitors of COS. Here, we describe that phloroglucinol, a natural phenolic compound having anti-COS properties, protects both rat RPE and mouse photoreceptor primary cultures from atRAL-induced cell death and reduces hydrogen peroxide (H2 O2 )-induced damage in RPE in a dose-dependent manner. Mechanistic analyses demonstrate that the protective effect encompasses decrease in atRAL-induced intracellular reactive oxygen species and free atRAL levels. Moreover, we show that phloroglucinol reacts with atRAL to form a chromene adduct which prevents bisretinoid A2E synthesis in vitro. Taken together, these data show that the protective effect of phloroglucinol correlates with its ability to trap atRAL and to prevent its further transformation into deleterious bisretinoids. Phloroglucinol might be a good basis to develop efficient therapeutic derivatives in the treatment of retinal macular diseases.
Assuntos
Citoproteção/efeitos dos fármacos , Floroglucinol/farmacologia , Células Fotorreceptoras de Vertebrados/metabolismo , Substâncias Protetoras/farmacologia , Epitélio Pigmentado da Retina/patologia , Retinaldeído/toxicidade , Retinoides/metabolismo , Animais , Benzopiranos/metabolismo , Morte Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Peróxido de Hidrogênio/toxicidade , Estresse Oxidativo , Espectroscopia de Prótons por Ressonância Magnética , Ratos Long-Evans , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/metabolismoRESUMO
Emixustat is a visual cycle modulator that has entered clinical trials as a treatment for age-related macular degeneration (AMD). This molecule has been proposed to inhibit the visual cycle isomerase RPE65, thereby slowing regeneration of 11-cis-retinal and reducing production of retinaldehyde condensation byproducts that may be involved in AMD pathology. Previously, we reported that all-trans-retinal (atRAL) is directly cytotoxic and that certain primary amine compounds that transiently sequester atRAL via Schiff base formation ameliorate retinal degeneration. Here, we have shown that emixustat stereoselectively inhibits RPE65 by direct active site binding. However, we detected the presence of emixustat-atRAL Schiff base conjugates, indicating that emixustat also acts as a retinal scavenger, which may contribute to its therapeutic effects. Using agents that lack either RPE65 inhibitory activity or the capacity to sequester atRAL, we assessed the relative importance of these 2 modes of action in protection against retinal phototoxicity in mice. The atRAL sequestrant QEA-B-001-NH2 conferred protection against phototoxicity without inhibiting RPE65, whereas an emixustat derivative incapable of atRAL sequestration was minimally protective, despite direct inhibition of RPE65. These data indicate that atRAL sequestration is an essential mechanism underlying the protective effects of emixustat and related compounds against retinal phototoxicity. Moreover, atRAL sequestration should be considered in the design of next-generation visual cycle modulators.
Assuntos
Éteres Fenílicos/farmacologia , Propanolaminas/farmacologia , Degeneração Retiniana/prevenção & controle , Transportadores de Cassetes de Ligação de ATP/deficiência , Transportadores de Cassetes de Ligação de ATP/genética , Oxirredutases do Álcool/deficiência , Oxirredutases do Álcool/genética , Animais , Domínio Catalítico , Bovinos , Cristalografia por Raios X , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/metabolismo , Sequestradores de Radicais Livres/farmacologia , Degeneração Macular/tratamento farmacológico , Degeneração Macular/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Moleculares , Éteres Fenílicos/química , Éteres Fenílicos/metabolismo , Propanolaminas/química , Propanolaminas/metabolismo , Degeneração Retiniana/metabolismo , Degeneração Retiniana/patologia , Retinaldeído/química , Retinaldeído/metabolismo , Retinaldeído/toxicidade , Bases de Schiff/metabolismo , Estereoisomerismo , cis-trans-Isomerases/antagonistas & inibidores , cis-trans-Isomerases/química , cis-trans-Isomerases/metabolismoRESUMO
Cellular events responsible for the initiation of major neurodegenerative disorders of the eye leading to blindness, including age-related macular degeneration, Stargardt and Best diseases, are poorly understood. Accumulation of vitamin A dimers, such as N-retinylidene-N-retinylethanolamine (A2E) in the retinal pigment epithelium (RPE), is one of the earliest measurable events preceding retinal degeneration. However, the extent to which these dimers contribute to tissue degeneration is not clear. To determine if A2E could trigger morphological changes associated with the degenerating RPE and subsequent cell death, we evaluated its toxicity to cultured human RPE cells (ARPE-19). We show that A2E triggered the accumulation of debris followed by a protracted death. A2E was up to ≈ 14-fold more toxic than its precursor, retinaldehyde. Measurements reveal that the concentration of A2E in the aged human eye could exceed the concentration of all other retinoids, opening the possibility of A2E-triggered cell death by several reported mechanisms. Findings suggest that accumulation of vitamin A dimers such as A2E in the human eye might be responsible for the formation of ubiquitous RPE debris, an early indication of retinal degeneration, and that preventing or reducing the accumulation of vitamin A dimers is a prudent strategy to prevent blindness.
Assuntos
Células Epiteliais/citologia , Degeneração Retiniana/metabolismo , Epitélio Pigmentado da Retina/citologia , Vitamina A/metabolismo , Idoso , Idoso de 80 Anos ou mais , Morte Celular , Células Cultivadas , Dimerização , Células Epiteliais/metabolismo , Humanos , Degeneração Retiniana/fisiopatologia , Epitélio Pigmentado da Retina/metabolismo , Retinaldeído/metabolismo , Retinaldeído/toxicidade , Vitamina A/química , Vitamina A/toxicidadeRESUMO
The current study investigates the cellular events which trigger activation of proapoptotic Bcl-2-associated × protein (Bax) in retinal cell death induced by all-trans-retinal (atRAL). Cellular events which activate Bax, such as DNA damage by oxidative stress and phosphorylation of p53, were evaluated by immunochemical and biochemical methods using ARPE-19 cells, 661 W cells, cultured neural retinas and a retinal degeneration model, Abca4(-/-)Rdh8(-/-) mice. atRAL-induced Bax activation in cultured neural retinas was examined by pharmacological and genetic methods. Other Bax-related cellular events were also evaluated by pharmacological and biochemical methods. Production of 8-OHdG, a DNA damage indicator, and the phosphorylation of p53 at Ser46 were detected prior to Bax activation in ARPE-19 cells incubated with atRAL. Light exposure to Abca4(-/-)Rdh8(-/-) mice also caused the above mentioned events in conditions of short term intense light exposure and regular room lighting conditions. Incubation with Bax inhibiting peptide and deletion of the Bax gene partially protected retinal cells from atRAL toxicity in cultured neural retina. Necrosis was demonstrated not to be the main pathway in atRAL mediated cell death. Bcl-2-interacting mediator and Bcl-2 expression levels were not altered by atRAL in vitro. atRAL-induced oxidative stress results in DNA damage leading to the activation of Bax by phosphorylated p53. This cascade is closely associated with an apoptotic cell death mechanism rather than necrosis.
Assuntos
Apoptose/efeitos dos fármacos , Dano ao DNA , Retina/patologia , Epitélio Pigmentado da Retina/patologia , Retinaldeído/toxicidade , Proteína X Associada a bcl-2/metabolismo , 8-Hidroxi-2'-Desoxiguanosina , Transportadores de Cassetes de Ligação de ATP/genética , Oxirredutases do Álcool/genética , Animais , Linhagem Celular , Colorimetria , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Modelos Animais de Doenças , Deleção de Genes , Humanos , Immunoblotting , Imuno-Histoquímica , L-Lactato Desidrogenase/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Técnicas de Cultura de Órgãos , Fosforilação , Retina/metabolismo , Degeneração Retiniana/metabolismo , Degeneração Retiniana/prevenção & controle , Epitélio Pigmentado da Retina/metabolismo , Tomografia de Coerência Óptica , Proteína Supressora de Tumor p53/metabolismoRESUMO
All-trans-retinal (AtRal) can accumulate in the retina as a result of excessive exposure to light. The purpose of this study was to compare cytotoxicity of AtRal and photodegraded AtRal (dAtRal) on cultured human retinal pigment epithelial cells in dark and upon exposure to visible light. AtRal was degraded by exposure to visible light. Cytotoxicity was monitored by imaging of cell morphology, propidium iodide staining of cells with permeable plasma membrane and measurements of reductive activity of cells. Generation of singlet oxygen photosensitized by AtRal and dAtRal was monitored by time-resolved measurements of characteristic singlet oxygen phosphorescence. Photodegradation of AtRal resulted in a decrease in absorption of visible light and accumulation of the degradation products with absorption maximum at â¼330 nm. Toxicity of dAtRal was concentration-dependent and was greater during irradiation with visible light than in dark. DAtRal was more cytotoxic than AtRal both in dark and during exposure to visible light. Photochemical properties of dAtRal indicate that it may be responsible for the maximum in the action spectra of retinal photodamage recorded in animals. In conclusion, photodegradation products of AtRal may impose a significant threat to the retina and therefore their roles in retinal pathology need to be explored.
Assuntos
Células Epiteliais/efeitos dos fármacos , Luz , Fotólise , Epitélio Pigmentado da Retina/efeitos dos fármacos , Retinaldeído/toxicidade , Linhagem Celular , Células Cultivadas , Dimetil Sulfóxido , Células Epiteliais/efeitos da radiação , Humanos , Oxigênio/metabolismo , Epitélio Pigmentado da Retina/citologia , Epitélio Pigmentado da Retina/efeitos da radiação , Retinaldeído/química , Retinaldeído/metabolismoRESUMO
The photophysical properties of all-trans-retinal (RAL) have been extensively studied because of the importance of the retinoids in the visual process. However, little information is available regarding the participation of RAL in photochemical transformations such as photoxidation. RAL is one of several native chromophores that have been suggested to act as photosensitizers of damage in the human retina, and this damage would likely occur through oxidative pathways. Time-resolved and steady state techniques have been used to examine the photoreactivity of RAL toward several suitable substrates. The lifetime of the RAL triplet excited state is observed to decrease with increasing concentration of the well-known electron and hydrogen atom donors, 2,3,5,6-tetramethyl-1,4-phenylenediamine (DAD), hydroquinone (HQ), methylhydroquinone (MHQ), 2,3-dimethylhydroquinone (DMHQ) and trimethylhydroquinone (TMHQ), although the bimolecular rate constants for the reaction are much less than that of diffusion controlled (2.9 x 10(7) M-1 s-1, 1.2 x 10(5) M-1 s-1, 1.2 x 10(5) M-1 s-1, 1.5 x 10(5) M-1 s-1 and 1.6 x 10(6) M-1 s-1, for DAD, HQ, MHQ, DMHQ and TMHQ, respectively). In the presence of the donors, new absorptions grow concomitant with the decay of the triplet excited state, and for DAD and TMHQ, the observed spectra are similar to the spectra of p-phenylenediamine and TMHQ radicals. Irradiation of RAL in argon-saturated methanol results in fairly efficient photobleaching of RAL and in the formation of two new compounds having absorption spectra that are shifted below 300 nm. Irradiation of RAL in argon-saturated acetonitrile also results in photobleaching of RAL, but the reaction proceeds at a slower rate.
Assuntos
Fármacos Fotossensibilizantes/toxicidade , Retinaldeído/toxicidade , Humanos , Técnicas In Vitro , Oxirredução , Fotoquímica , Fármacos Fotossensibilizantes/efeitos da radiação , Retina/lesões , Retina/efeitos da radiação , Retinaldeído/efeitos da radiação , EspectrofotometriaRESUMO
Recent intervention studies revealed that beta-carotene supplement to smokers resulted in a higher incidence of lung cancer. However, the causal mechanisms remain to be clarified. We reported here that vitamin A (retinol) and its derivative (retinal) caused cellular DNA cleavage detected by pulsed field gel electrophoresis. Retinol and retinal significantly induced 8-oxo-7,8-dihydro-2'-deoxyguanosine formation in HL-60 cells but not in H(2)O(2)-resistant HP100 cells, suggesting the involvement of H(2)O(2) in cellular DNA damage. Experiments using (32)P-labeled isolated DNA demonstrated that retinol and retinal caused Cu(II)-mediated DNA damage, which was inhibited by catalase. UV-visible spectroscopic and electron spin resonance-trapping studies revealed the generation of superoxide and carbon-centered radicals, respectively. The superoxide generation during autoxidation of retinoids was significantly correlated with the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine, although the yield of carbon-centered radicals was not necessarily related to the intensity of DNA damage. These findings suggest that superoxide generated by autoxidation of retinoids was dismutated to H(2)O(2), which was responsible for DNA damage in the presence of endogenous metals. Retinol and retinal have prooxidant abilities, which might lead to carcinogenesis of the supplements of beta-carotene.
Assuntos
Dano ao DNA , Retinaldeído/toxicidade , Superóxidos/metabolismo , Vitamina A/toxicidade , 8-Hidroxi-2'-Desoxiguanosina , Animais , Bovinos , Linhagem Celular , Enzimas de Restrição do DNA/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Relação Dose-Resposta a Droga , Espectroscopia de Ressonância de Spin Eletrônica , Eletroforese em Gel de Campo Pulsado , Sequestradores de Radicais Livres/farmacologia , Radicais Livres/metabolismo , Genes p53/genética , Genes ras/genética , Células HL-60 , Humanos , Peróxido de Hidrogênio/metabolismo , Fenantrolinas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Especificidade por Substrato , Timo/metabolismo , Fatores de TempoRESUMO
BACKGROUND: We had hypothesised that retinaldehyde (RAL) should be an interesting precursor for topical use. AIM: We review our observations about its biological activities. METHODS: We performed pilot studies to explore its biological effects and tolerability in human skin and compared the effects of topical RAL to that of all-trans-retinoic acid (RA) in the mouse tail test. RESULTS: The biological activities of RAL were found to be qualitatively identical to that of RA: (i) induction of cellular RA-binding protein type 2 mRNA and protein, (ii) increase in epidermal proliferation (increase in DNA synthesis, epidermal thickness, induction of 50-kD keratin mRNA and reduction in 70-kD keratin mRNA), and (iii) metaplastic effects (induction of orthokeratosis, reduction of 65-kD keratin mRNA, increase in filaggrin and loricrin mRNAs). When associated with RAL, citral (known for its capacity to inhibit the oxidation of retinol to RA in epidermis) counteracted the effects induced by RAL indicating that RAL exerts biological activities through transformation to RA. Hypothesizing that keratinocytes would metabolize 9-cis-RAL to 9-cis-RA, we compared the biological effects induced by topical 9-cis-RAL and found that hyperplastic and metaplastic responses were lower than those induced by all-trans-RAL or all-trans-RA at similar concentrations. This suggests that 9-cis-RAL has no advantage over all-trans-RAL for specific delivery of natural retinoids into the skin. As in clinical studies conducted in human skin, we also found topical RAL less irritant than RA. CONCLUSION: These studies indicate that topical RAL has biological activity and is well tolerated.
Assuntos
Monoterpenos , Retinaldeído/farmacologia , Pele/efeitos dos fármacos , Monoterpenos Acíclicos , Administração Tópica , Animais , Relação Dose-Resposta a Droga , Proteínas Filagrinas , Humanos , Queratinas/metabolismo , Ceratolíticos/administração & dosagem , Ceratolíticos/farmacologia , Camundongos , Peroxidase/metabolismo , Retinaldeído/administração & dosagem , Retinaldeído/toxicidade , Pele/metabolismo , Terpenos/farmacologia , Tretinoína/administração & dosagem , Tretinoína/farmacologia , Tretinoína/toxicidadeRESUMO
Recently, the temporal and spatial distribution patterns of two established, endogenous retinoid receptor ligands, 9-cis-retinoic acid and all-trans-retinoic acid and various precursor retinoids were described in Xenopus embryos during early development (Creech Kraft et al., Proc. Natl. Acad. Sci. U.S.A. 1994; Biochem. J. 1994). Each of these two receptor ligands is a metabolite of vitamin A (all-trans-retinol), and each is also a potent dysmorphogen in Xenopus embryos as well as in embryos of several other vertebrate species. This study demonstrates early embryonic metabolism of exogenous all-trans-retinol, 9-cis-retinol, all-trans-retinal, and 9-cis-retinal to 9-cis-retinoic acid, all-trans-retinoic acid, and other metabolites in Xenopus embryos during neurulation, a specific stage of development that spans a time period of approximately 8 hr. Our results demonstrate that the Xenopus embryo provides a suitable model system for studying the embryonic bioconversion of retinoids and dysmorphogenic effects within a single time window of development.