Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 46
Filter
1.
Journal of Experimental Hematology ; (6): 1065-1070, 2021.
Article in Chinese | WPRIM | ID: wpr-888519

ABSTRACT

OBJECTIVE@#To investigate the expression of peptidylarginine deiminase 4 (PADI4) during the process of differentiation into granulocyte of NB4 cells induced by all-trans-retinoic acid (ATRA) and whether PADI4 is involved in the inflammatory cytokines expression.@*METHODS@#Granulocyte differentiation model of NB4 cells induced by ATRA was established. The cell morphology changes were observed by Wright-Giemsa staining. The expression of cell differentiation marker CD11b was analyzed by flow cytometry. The mRNA and protein expression of PADI4 was detected by RT-PCR and Western blot, respectively. The expression of tumor necrosis factor (TNF) α and interleukin (IL) 1β was analyzed by ELISA, and also examined with the knockdown of PADI4 expression by siRNA.@*RESULTS@#After NB4 cells induced by ATRA, the cytoplasm increased and the ratio of nuclear to cytoplasmic was reduced. Nuclear dented, and rod-shaped nucleus, lobulated phenomenon increased (P<0.05). Flow cytometry analysis results showed that the cell surface molecule CD11b expression increased (P<0.01). RT-PCR and Western blot showed the expression of PADI4 increased at both transcriptional and translational levels during the process of the differentiation. ELISA showed TNF-α and IL-1β secretion increased in differentiated macrophages, while they could be inhibited by PADI4-specific siRNA.@*CONCLUSION@#During the differentiation into granulocyte of NB4 cells induced by ATRA, PADI4 expression increased. Furthermore, PADI4 appeared to play a critical role in inflammatory cytokines secretion.


Subject(s)
Cell Differentiation , Cell Line, Tumor , Cytokines/metabolism , Granulocytes , Humans , Leukemia, Promyelocytic, Acute , Protein-Arginine Deiminase Type 4/metabolism , Tretinoin/pharmacology
2.
Acta cir. bras ; 35(1): e202000106, 2020. graf
Article in English | LILACS | ID: biblio-1088526

ABSTRACT

Abstract Purpose To explore the role of all-trans retinoic acid (ATRA) in renal ischemia/reperfusion injury of diabetic rats. Methods Sixty adult male rats were randomly divided into 6 groups, including sham group (S group), ischemia-reperfusion group (I/R group), ischemia-reperfusion+ATRA group (A group), diabetic group (D group), diabetic ischemia-reperfusion group (DI/R group), diabetic ischemia-reperfusion +ATRA group (DA group). The levels of creatinine (Cr), cystatin C (Cys-C) and β2-microglobulin (β2-MG) were measured. Morphology of renal tissue was observed under light microscope. Results DJ-1, Nrf2, HO-1 and caspase-3 were detected by western blot. DJ-1, Nrf2, HO-1 and caspase-3 in I/R group, D group and DI/R group was higher than that in S group. Compared with I/R group, Nrf2 and HO-1 in A group was decreased, but caspase-3 was increased. However, Nrf2 in DA group was higher than that in DI/R group, HO-1 and caspase-3 in DA group were lower than that in DI/R group. Compared with group S, Cr, Cys-C and β2-MG in I/R group, A group, D group, and DI/R group were higher. Whereas the levels of Cr, Cys-C, β2-MG and renal injury score in DA group were lower than those in DI/R group. Conclusion ATRA has a protective effect on renal ischemia-reperfusion injury in diabetic rats, maybe relating to DJ/Nrf2 pathway.


Subject(s)
Animals , Male , Rats , Tretinoin/therapeutic use , Reperfusion Injury/prevention & control , Diabetes Mellitus, Experimental/chemically induced , NF-E2-Related Factor 2/therapeutic use , Kidney/drug effects , Tretinoin/pharmacology , Reperfusion Injury/pathology , Streptozocin , Disease Models, Animal , Drug Evaluation, Preclinical , NF-E2-Related Factor 2/pharmacology , Kidney/pathology
3.
J. appl. oral sci ; 27: e20180317, 2019. tab, graf
Article in English | LILACS, BBO | ID: biblio-984571

ABSTRACT

Abstract Bone morphogenetic protein type 2 (BMP-2) and retinoic acid (RA) are osteoinductive factors that stimulate endogenous mechanisms of bone repair which can be applied on management of osseous defects in oral and maxillofacial fields. Objective Considering the different results of RA on osteogenesis and its possible use to substitute/potency BMP-2 effects, this study evaluated the outcomes of BMP-2, RA, and BMP-2+RA treatments on in vitro osteogenic differentiation of human adipose-derived stem cells (ASCs) and the signaling pathway(s) involved. Material and Methods ASCs were treated every other day with basic osteogenic medium (OM) alone or supplemented with BMP-2, RA, or BMP-2+RA. Alkaline phosphatase (ALP) activity was determined using the r-nitrophenol method. Extracellular matrix mineralization was evaluated using von Kossa staining and calcium quantification. Expression of osteonectin and osteocalcin mRNA were determined using qPCR. Smad1, Smad4, phosphorylated Smad1/5/8, BMP-4, and BMP-7 proteins expressions were analyzed using western blotting. Signaling pathway was evaluated using the IPA® software. Results RA promoted the highest ALP activity at days 7, 14, 21, and 28, in comparison to BMP-2 and BMP-2+RA. BMP-2+RA best stimulated phosphorylated Smad1/5/8 protein expression at day 7 and Smad4 expression at days 7, 14, 21, and 28. Osteocalcin and osteonectin mRNA expressions were best stimulated by BMP-2+RA at day 7. Matrix mineralization was most improved by BMP-2+RA at days 12 and 32. Additionally, BMP-2+RA promoted the highest BMP signaling pathway activation at days 7 and 14, and demonstrated more activation of differentiation of bone-forming cells than OM alone. Conclusions In summary, RA increased the effect of BMP-2 on osteogenic differentiation of human ASCs.


Subject(s)
Humans , Osteogenesis/drug effects , Tretinoin/pharmacology , Cell Differentiation/drug effects , Bone Morphogenetic Protein 2/drug effects , Mesenchymal Stem Cells/drug effects , Osteoblasts/drug effects , Osteogenesis/physiology , Reference Values , Time Factors , Osteocalcin/analysis , Osteocalcin/drug effects , Osteonectin/analysis , Osteonectin/drug effects , Cell Differentiation/physiology , Cells, Cultured , Blotting, Western , Reproducibility of Results , Analysis of Variance , Alkaline Phosphatase/analysis , Alkaline Phosphatase/adverse effects , Bone Morphogenetic Protein 2/metabolism , Mesenchymal Stem Cells/metabolism
4.
Electron. j. biotechnol ; 34: 43-50, july. 2018. tab, graf, ilus
Article in English | LILACS | ID: biblio-1045999

ABSTRACT

Background: All-trans retinoic acid (ATRA), a vitamin A-derived active metabolite, exerts important functions in hair biology. Previous studies indicated that excess ATRA hampered hair follicle morphogenesis and cyclic regeneration in adulthood, but other studies stated that ATRA promoted hair growth. Dermal papilla (DP), a cluster of specialized fibroblasts, plays pivotal roles in controlling development and regeneration of hair follicle. Several lines of evidence indicated that DP might be the target cells of ATRA in the hair follicle. To confirm this hypothesis, the present study was performed to explore the biological effects of ATRA on goat dermal papilla cells (DPCs) and clarify the roles of ATRA in hair biology. Results: Our experimental results indicated that key signaling transducers of ATRA were dynamically expressed in distinct stages of goat cashmere growth cycle, and high-dose ATRA treatment (10-5 M) significantly impaired the viability of goat DPCs and lowered the ratio of proliferating cells. Otherwise, goat DPCs were stimulated to enter apoptosis and their cell cycle progression was severely blocked by ATRA. Moreover, the expression of fibroblast growth factor 7 (Fgf7), one of the potent hair growth stimulators secreted by DPCs, was transcriptionally repressed following ATRA treatment. Conclusion: DPCs are the targets of ATRA in the hair follicle, and ATRA negatively regulates hair growth by the targeted suppression of cell viability and growth factor expression of goat DPCs. Through these observations, we offer a new mechanistic insight into the roles of ATRA in hair biology.


Subject(s)
Animals , Tretinoin/pharmacology , Goats , Hair Follicle/drug effects , Regeneration , In Vitro Techniques , Immunohistochemistry , Receptors, Retinoic Acid , Hair Follicle/cytology , Hair Follicle/growth & development , Cell Proliferation/drug effects , Fibroblast Growth Factor 7/genetics , Real-Time Polymerase Chain Reaction
5.
An. bras. dermatol ; 92(2): 191-195, Mar.-Apr. 2017. graf
Article in English | LILACS | ID: biblio-838051

ABSTRACT

Abstract: Background: Isotretinoin is a synthetic analog of vitamin A. Recent studies support a role for retinoic acid in the recovery of olfactory function following injury in mice. Objective: This study aimed at determining the effect of isotretinoin on olfactory function in patients who have acne and are otherwise healthy. Methods: Forty-five patients (aged 25-40 years) with acne were included in the study. All patients underwent a rhinological examination. Olfactory function was assessed by the Sniffin' Sticks Test. The test was assessed at baseline and in the third month of isotretinoin treatment. Results: Isotretinoin improved the performance of patients in the olfactory test. The SST score increased from 8.7±1.09 to 9.5±1.19 (p<0.001), prevalence of hyposmia decreased from 40% to 24% and normosmia increased from 60% to 75% (p=0.059). The percentage of patients whose olfactory function was categorized as "good" increased from 6% to 21.3%. This increase was statistically significant (p<0.05). Study limitations: Absence of a control group is one of the limitations of this study. Also, we did not evaluate patients with smell test after stopping isotretinoin treatment. Conclusion: We examined the effect of systemic isotretinoin on olfactory function. It can be concluded from the present investigation that isotretinoin therapy improves the sense of smell.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Young Adult , Smell/drug effects , Tretinoin/therapeutic use , Isotretinoin/therapeutic use , Acne Vulgaris/drug therapy , Tretinoin/pharmacology , Isotretinoin/pharmacology , Prospective Studies
6.
Braz. j. med. biol. res ; 50(4): e5561, 2017. graf
Article in English | LILACS | ID: biblio-839280

ABSTRACT

The aim of this study was to investigate whether exogenous retinoic acid (RA) can upregulate the mRNA and protein expression of growth-associated protein 43 (GAP-43), thereby promoting brain functional recovery in a rat distal middle cerebral artery occlusion (MCAO) model of ischemia. A total of 216 male Sprague Dawley rats weighing 300–320 g were divided into 3 groups: sham-operated group, MCAO+vehicle group and MCAO+RA group. Focal cortical infarction was induced with a distal MCAO model. The expression of GAP-43 mRNA and protein in the ipsilateral perifocal region was assessed using qPCR and immunocytochemistry at 1, 3, 7, 14, 21, and 28 days after distal MCAO. In addition, an intraperitoneal injection of RA was given 12 h before MCAO and continued every day until the animal was sacrificed. Following ischemia, the expression of GAP-43 first increased considerably and then decreased. Administration of RA reduced infarction volume, promoted neurological functional recovery and upregulated expression of GAP-43. Administration of RA can ameliorate neuronal damage and promote nerve regeneration by upregulating the expression of GAP-43 in the perifocal region after distal MCAO.


Subject(s)
Animals , Male , GAP-43 Protein/metabolism , Gene Expression/drug effects , Infarction, Middle Cerebral Artery/prevention & control , Neuroprotective Agents/pharmacology , Tretinoin/pharmacology , Up-Regulation/drug effects , Brain Ischemia/prevention & control , GAP-43 Protein/genetics , Immunohistochemistry , Infarction, Middle Cerebral Artery/metabolism , Infarction, Middle Cerebral Artery/pathology , Random Allocation , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Time Factors
7.
Article in English | WPRIM | ID: wpr-141165

ABSTRACT

Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1alpha, 25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD3. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD3 increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD3-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.


Subject(s)
Antibodies, Bacterial/immunology , CD11c Antigen/metabolism , Lipopolysaccharide Receptors/metabolism , CD18 Antigens/metabolism , Apoptosis/immunology , Biological Assay , Cell Differentiation , Cell Line, Tumor , Cholecalciferol/pharmacology , Dimethylformamide/pharmacology , Flow Cytometry , HL-60 Cells , Humans , Phagocytosis/immunology , Pneumococcal Vaccines/immunology , Receptors, IgG/metabolism , Receptors, Immunologic/biosynthesis , Respiratory Burst/immunology , Streptococcus pneumoniae/immunology , Tretinoin/pharmacology
8.
Article in English | WPRIM | ID: wpr-141164

ABSTRACT

Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1alpha, 25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD3. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD3 increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD3-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.


Subject(s)
Antibodies, Bacterial/immunology , CD11c Antigen/metabolism , Lipopolysaccharide Receptors/metabolism , CD18 Antigens/metabolism , Apoptosis/immunology , Biological Assay , Cell Differentiation , Cell Line, Tumor , Cholecalciferol/pharmacology , Dimethylformamide/pharmacology , Flow Cytometry , HL-60 Cells , Humans , Phagocytosis/immunology , Pneumococcal Vaccines/immunology , Receptors, IgG/metabolism , Receptors, Immunologic/biosynthesis , Respiratory Burst/immunology , Streptococcus pneumoniae/immunology , Tretinoin/pharmacology
9.
Yonsei Medical Journal ; : 1597-1603, 2015.
Article in English | WPRIM | ID: wpr-177064

ABSTRACT

PURPOSE: The aim of this study was to examine the effects of all-trans retinoic acid (ATRA) on diabetic nephropathy. MATERIALS AND METHODS: We measured amounts of urinary albumin excretion (UAE) after administrating ATRA to Otsuka Long-Evans Tokushima Fatty (OLETF) rats. In order to understand the mechanism of action for ATRA, we administrated ATRA to examine its inhibitory action on the production of transforming growth factor-beta1 (TGF-beta1), protein kinase C (PKC), and reactive oxidative stress (ROS) in cultured rat mesangial cells (RMCs). RESULTS: After 16 weeks of treatment, UAE was lower in the ATRA-treated OLETF rats than in the non-treated OLETF rats (0.07+/-0.03 mg/mgCr vs. 0.17+/-0.15 mg/mgCr, p<0.01). After incubation of RMCs in media containing 30 or 5 mM of glucose, treatment with ATRA showed time- and dose-dependent decreases in TGF-beta1 levels and ROS. Moreover, ATRA treatment showed a dose-dependent decrease in PKC expression. CONCLUSION: ATRA treatment suppressed UAE and TGF-beta1 synthesis, which was mediated by significant reductions in PKC activity and ROS production. Our results suggest that ATRA has a potential therapeutic role for diabetic nephropathy.


Subject(s)
Animals , Diabetes Mellitus, Type 2/complications , Diabetic Nephropathies/complications , Mesangial Cells/metabolism , Oxidative Stress/drug effects , Rats , Rats, Inbred OLETF , Reactive Oxygen Species/metabolism , Transforming Growth Factor beta1/analysis , Tretinoin/pharmacology
10.
Journal of Gorgan University of Medical Sciences. 2015; 17 (3): 46-54
in Persian | IMEMR | ID: emr-173783

ABSTRACT

Background and Objective: With respect to the antioxidant role of melatonin and retinoic acid, it seems to be effective both in the maturation and embryonic development. This study was done to investigate the effect of combination of melatonin and All-Trans retinoic acid [RA] on maturation, fertilization and embryonic development of immature mouse oocytes


Methods: In this experimental study, cumulus - oocyte complex [COCs] were recovered from 4-6 week old female mice NMRI and were divided into 6 maturation medium groups including control, sham, experiment 1[melatonin 100 nM, 1 and 2 microM], experiment 2 [retinoic acid 1, 2, 4, 6 microM], experiment 3 [melatonin 2 microM+RA 4 microM], experiment 4 [Mel 100nM + retinoic acid 4 microM]. The maturation rate was recorded after 24 hours of culture in a humidified atmosphere of 5% CO2 at 37°C. The matured oocytes were fertilized with sperm. Fertilization and embryonic development rates to the blastocyst stage were recorded


Results: Maturation rate in the control and sham groups were 50.6% and 49.4%, respectively. Maturation rate were 54.3%, 54.8%, 59.9% in melatonin group with concentrations of 100 nM, 1 and 2 microM, respectively. Maturation rate were 51.6%, 51%, 59% and 49.6% in t-RA group with concentrations of 1, 2, 4, 6 microM. Maturation rate were 60.4% and 54.2% in the experiment 3 and 4 groups, respectively. The maturation rates in the melatonin 2 microM, retinoic acid 4 microM and experiment 3 significantly increased in compare to control [P<0.05]. The embryonic development rate in the melatonin with 100nM concentration and 4 microM of retinoic acid increased significantly compared to controls [P<0.05]. Although, embryonic development rate in experiment 3 was higher than control, but lower in compare to melatonin 100 nM and the retinoic acid 4 microM. The embryonic development rate in experiment 4 significantly increased in compare to control [P<0.05]


Conclusion: Combination of melatonin and All-Trans retinoic acid in medium culture increase maturation rate and improved embryonic development in dose dependent manner


Subject(s)
Animals, Laboratory , Tretinoin/pharmacology , Oocytes , Fertilization , In Vitro Oocyte Maturation Techniques , Embryonic Development , Mice
11.
Int. j. morphol ; 32(4): 1449-1456, Dec. 2014. ilus
Article in Spanish | LILACS | ID: lil-734697

ABSTRACT

El déficit y exceso de vitamina A provoca malformaciones congénitas que afectan distintos órganos y sistemas. El objetivo de este estudio fue determinar el efecto que causa la administración de ácido retinoico a distintas dosis sobre la morfogénesis ósea del esqueleto axial en embriones de ratón Mus musculus. Mediante aleatorización simple se distribuyeron hembras recién preñadas en 4 categorías: A, B, C y D. El día 8 post fecundación (p.f), se administró 40 mg/kg de peso de ácido retinoico al grupo A, 20 mg/kg de peso de esta solución al grupo B, 1 ml/kg de peso de dimetil sulfóxido al grupo C, y el grupo D es grupo control. El día 17 de la gestación las hembras y sus fetos fueron anestesiadas y eutanasiadas con sobredosis de pentotal sódico intraperitoneal. Los fetos de cada camada fueron procesados mediante diafanización y tinción con azul de Alcian para destacar cartílago hialino y alizarina para observar tejido óseo. Los resultados se expresaron en porcentajes de malformaciones en los siguientes tres segmentos: 1) cráneo-columna cervical, 2) segmento torácico y abdominal y 3) cintura pélvica, considerándose un 100% cuando la totalidad de los elementos óseos se encontraban comprometidos. Se utilizó la prueba de Fisher para la comparación de frecuencias de malformaciones y se consideró estadísticamente significativo cuando p<0,05. En el grupo A se evidenciaron malformaciones mayores como ausencia de huesos frontales y parietales, exencefalia, defectos en el número de vértebras, y fusiones de costillas; y en el grupo B se observaron malformaciones menores como alteraciones numéricas y fusiones de costillas, existiendo diferencias significativas entre ambos grupos. En los grupos C y D no se consignaron malformaciones. El ácido retinoico administrado intraperitonealmente el dìa 8 p.f en dosis de 40 y 20 mg/kg de peso se comporta como un teratógeno en los embriones de ratón, existiendo además diferencias significativas entre las malformaciones generadas por ambas dosis de ácido retinoico. La primera concentración afecta los huesos de los tres segmentos estudiados (cráneo-cervical, toracoabdominal, y pélvico) y la segunda concentración sólo afecta a dos segmentos (cráneo-cervical y toracoabdominal). Ambos tratamientos afectan los segmentos en una gradiente céfalo caudal, independiente del origen embrionario de las estructuras. Esto se debería a que los cambios en las gradientes de ácido retinoico alteran el comportamiento de células de la cresta neural craneal y el orden de la expresión de genes Hox.


The deficit and excess of vitamin A causes birth defects affecting different organ systems. The objectives of this study are to determine the effect caused by the administration of different doses of retinoic acid on bone morphogenesis of the axial skeleton in embryonic mouse Mus musculus. By simple randomization newly pregnant females were distributed into 4 categories: A, B, C and D. On day 8 post fertilization, 40 mg/kg was administered by weight of retinoic acid to the group A, 20 mg/kg body weight of the group B solution 1 ml/kg body weight of dimethyl sulfoxide and group C. Group D is the control group. On day 17 of gestation the females and their fetuses were anesthetized and euthanized with an overdose of intraperitoneal sodium pentothal. Fetuses from each litter were processed using diaphanization and Alcian blue staining to hyaline cartilage and alizarin to observe bone tissue. The results are expressed as percentages of malformations in the following three segments: 1) cranio-cervical spine, 2) thoracic and abdominal segment and 3) pelvic segment, considering 100% when all the bony elements were compromised. Fisher's exact test for comparison of frequencies of malformations was used and considered statistically significant when p<0.05. In group A, major malformations and defects were evident in the indemnity of the cranial vault, exencephaly, defects in the number of vertebrae, and fusion of ribs. In group B minor malformations as numerical alterations and rib fusions were observed. Significant differences were found between both groups. In groups C and D no malformations were recorded. Retinoic acid administered intraperitoneally at doses of 40 and 20 mg/kg acts as a teratogen in mouse embryos. There are significant differences between the defects induced by concentrations of 40 mg/k and 20 mg/k of retinoic acid. Both concentrations affect the bones of the three segments studied (cranio cervical, thoraco-abdominal and pelvic) in a cephalo caudal gradient, independent of the embryonic origin of the structures. Changes in retinoic acid concentration alter the behavior of cranial neural crest and changing the order of the HOX gene expression in the axial skeleton.


Subject(s)
Animals , Male , Female , Mice , Tretinoin/administration & dosage , Abnormalities, Multiple/chemically induced , Bone Development/drug effects , Embryo, Mammalian/drug effects , Teratogens , Tretinoin/pharmacology , Embryonic Development/drug effects
12.
An. bras. dermatol ; 88(6): 930-936, Nov-Dec/2013. graf
Article in English | LILACS | ID: lil-699007

ABSTRACT

BACKGROUND: The sum of environmental and genetic factors affects the appearance and function of the skin as it ages. The identification of molecular changes that take place during skin aging provides biomarkers and possible targets for therapeutic intervention. Retinoic acid in different formulations has emerged as an alternative to prevent and repair age-related skin damage. OBJECTIVES: To understand the effects of different retinoid formulations on the expression of genes associated with biological processes that undergo changes during skin aging. METHODS: Ex-vivo skin samples were treated topically with different retinoid formulations. The modulation of biological processes associated with skin aging was measured by Reverse Transcription quantitative PCR (RT-qPCR). RESULTS: A formulation containing microencapsulated retinol and a blend of active ingredients prepared as a triple nanoemulsion provided the best results for the modulation of biological, process-related genes that are usually affected during skin aging. CONCLUSION: This association proved to be therapeutically more effective than tretinoin or microencapsulated retinol used singly. .


FUNDAMENTOS: A soma de fatores genéticos e ambientais afeta a aparência e a funcionalidade da pele ao longo do envelhecimento. O conhecimento a respeito das mudanças moleculares durante o envelhecimento fornece biomarcadores e possíveis alvos para intervenções terapêuticas. O ácido retinoico em diferentes formulações surgiu como uma alternativa para prevenir e reparar os danos da pele associados ao envelhecimento. OBJETIVOS: Avaliar comparativamente os efeitos de diferentes formulações contendo retinoides na expressão de genes associados a processos biológicos que são alterados com o envelhecimento da pele. MÉTODOS: Peles ex vivo foram topicamente tratadas com diferentes retinoides, micro e nanoencapsulados. A modulação dos processos biológicos associados ao envelhecimento da pele foi medida por PCR quantitativa, precedida de transcrição reversa (RT-qPCR). RESULTADOS: A formulação contendo uma mistura de princípios ativos incorporados em uma tripla nanoemulsão e retinol microencapsulado apresentou os melhores resultados de modulação de genes relacionados a processos biológicos que são normalmente alterados durante o envelhecimento da pele. CONCLUSÃO: Essa associação demonstrou uma maior eficácia terapêutica quando comparada ao uso isolado de tretinoína ou retinol microencapsulado. .


Subject(s)
Humans , Keratolytic Agents/therapeutic use , Skin Aging/drug effects , Skin/drug effects , Tretinoin/therapeutic use , Administration, Topical , Analysis of Variance , Biological Phenomena/drug effects , Drug Synergism , Emulsions , Gene Expression , Keratolytic Agents/pharmacology , Nanomedicine , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Skin Aging/genetics , Tretinoin/pharmacology
13.
Braz. j. med. biol. res ; 45(8): 721-729, Aug. 2012. ilus, tab
Article in English | LILACS | ID: lil-643658

ABSTRACT

Hepatocellular carcinoma (HCC) is the third highest cause of cancer death worldwide. In general, the disease is diagnosed at an advanced stage when potentially curative therapies are no longer feasible. For this reason, it is very important to develop new therapeutic approaches. Retinoic acid (RA) is a natural derivative of vitamin A that regulates important biological processes including cell proliferation and differentiation. In vitro studies have shown that RA is effective in inhibiting growth of HCC cells; however, responsiveness to treatment varies among different HCC cell lines. The objective of the present study was to determine if the combined use of RA (0.1 µM) and cAMP (1 mM), an important second messenger, improves the responsiveness of HCC cells to RA treatment. We evaluated the proliferative behavior of an HCC cell line (HTC) and the expression profile of genes related to cancer signaling pathway (ERK and GSK-3β) and liver differentiation (E-cadherin, connexin 26 (Cx26), and Cx32). RA and cAMP were effective in inhibiting the proliferation of HTC cells independently of combined use. However, when a mixture of RA and cAMP was used, the signals concerning the degree of cell differentiation were increased. As demonstrated by Western blot, the treatment increased E-cadherin, Cx26, Cx32 and Ser9-GSK-3β (inactive form) expression while the expression of Cx43, Tyr216-GSK-3β (active form) and phosphorylated ERK decreased. Furthermore, telomerase activity was inhibited along treatment. Taken together, the results showed that the combined use of RA and cAMP is more effective in inducing differentiation of HTC cells.


Subject(s)
Animals , Rats , Carcinoma, Hepatocellular/pathology , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cyclic AMP/pharmacology , Liver Neoplasms/pathology , Tretinoin/pharmacology , Cell Line, Tumor , Carcinoma, Hepatocellular/metabolism , Drug Combinations , Enzyme-Linked Immunosorbent Assay , Immunoblotting , Liver Neoplasms/metabolism , Microscopy, Confocal , Mitotic Index , Polymerase Chain Reaction
14.
Iranian Journal of Allergy, Asthma and Immunology. 2011; 10 (4): 243-249
in English | IMEMR | ID: emr-118121

ABSTRACT

All-trans retinoic acid [ATRA], as an active metabolite of vitamin A, has been shown to affect immune cells. This study was performed to evaluate the effect of ATRA on viability, proliferation, activation and lineage-specific transcription factors of CD4+ T cells. CD4+ T cells were separated from heparinized blood of healthy donors and were cultured in conditions, some with, some without ATRA. Viability was assessed by PI flowcytometry and proliferation was measured by MTT assay. CD69 expression was determined by flowcytometry as a measure of cell activation. Lineage-specific transcription factors [FOXP3, RORgammat and T-bet] were examined by intracellular staining and flowcytometry. High doses of ATRA [0.1-1 mM] caused extensive cell death in both PBMCs and CD4+ T cells. Doses of ATRA equal to or lower than 10 microM did not adversely affect cell viability and proliferation in comparison to culture medium without ATRA. Doses of ATRA between 10 microM and InM significantly increased cell activation when compared to culture medium without ATRA. ATRA could increase FOXP3+ and also FOXP3+RORgammat+ T cells while it decreased RORgammat+ and T-bet+ T cells. This study showed that doses of ATRA up to 10 microM are safe when using with CD4+ T cells in terms of cell viability, proliferation and activation. We could also show that ATRA diverts the human immune response in neutral conditions [without adding polarizing cytokines] by increasing FOXP3+ cells and decreasing RORgammat+ cells. ATRA could be regarded as a potential therapy in inflammatory conditions and autoimmunities


Subject(s)
Humans , Tretinoin/pharmacology , CD4-Positive T-Lymphocytes/immunology , Dose-Response Relationship, Drug , Flow Cytometry , Lymphocyte Activation/drug effects , Cell Survival , Cell Lineage , T-Box Domain Proteins/analysis , Forkhead Transcription Factors/analysis
15.
Braz. j. pharm. sci ; 45(1): 177-183, jan.-mar. 2009. graf, tab
Article in Portuguese | LILACS | ID: lil-525785

ABSTRACT

O ácido retinóico (AR) tem sido utilizado para o tratamento de acne severa, rugas, estrias e celulite, no entanto, provoca irritação na pele e sofre rápida degradação quando exposto à luz e ao calor. Métodos analíticos rápidos para quantificação do AR são, portanto, necessários para ensaios de cinética de liberação in vitro. Nesse contexto, o objetivo deste trabalho foi desenvolver e validar um método rápido e sensível para o doseamento do AR em microcápsulas de alginato/quitosana contendo óleo de babaçu dispersas em gel natrosol® por cromatografia líquida de alta eficiência associada à espectroscopia UV e aplicá-lo na avaliação do perfil de liberação in vitro dessas formulações. As análises foram realizadas em modo isocrático utilizando coluna C18 de fase reversa 150 x 4,6 mm (5 μm) com detecção a 350 nm. A fase móvel foi constituída de metanol e ácido acético 1 por cento (85:15 v/v) com vazão de 1,8 mL/minuto. A faixa de linearidade do método foi de 0,5 a 60 μg/mL (r² = 0,999). O método validado mostrou-se sensível, específico, exato, preciso, de baixo custo e o tempo de retenção do AR foi de 5,8 ± 0,4 minutos sendo, desta forma, mais rápido do que os relatados na literatura.


Retinoic acid (RA) has been used in the treatment of severe acne, wrinkles and cellulite. However, it induces skin irritation and rapidly suffers degradation under light and high temperate exposure. Rapid analytical methods to quantify retinoic acid are therefore mandatory for in vitro drug release studies. In this framework, the aim of this study was to develop and validate a rapid and responsive method to quantify the RA in microcapsules of chitosan and alginate containing babassu oil dispersed in natrosol® hydrogel using high performance liquid chromatography (HPLC). Furthermore this method was used to quantify in vitro release kinetics of RA from microcapsules. The analyses have been carried through an isocratic HPLC-UV method using a reversed phase 150 x 4.6 mm C18 (5μm) column, a mobile phase constituted of methanol and 1 percent acetic acid (85:15) at a flow rate of 1.8 mL/min and detection at 350 nm. The linearity range was 0.5-60 μg/mL (r² = 0.999). The validated HPLC-UV method was responsive, specific, accurate, precise, and economic and the RA retention time was 5.8 ± 0.4 minutes, being therefore, faster than that previously reported.


Subject(s)
Alginates , Chitosan , Chromatography, Liquid/instrumentation , Tretinoin/pharmacology , Tissue Extracts/analysis , Pharmaceutical Preparations/analysis , Reproducibility of Results/methods
16.
Pakistan Journal of Pharmaceutical Sciences. 2009; 22 (1): 23-26
in English | IMEMR | ID: emr-92318

ABSTRACT

All-trans retinoic acid [ATRA] has beneficial and teratogenicity effects when used in a variety conditions. The objectives of this study were to determine the effects of ATRA on the Progenitors of red blood cell and platelets in rat's embryo. Single oral dose [100 mg/kg] of ATRA was administered to rat on gestation day [GD] 10 and fetuses were observed on GD 18 and compared with untreated group. In the experimental embryos of GD 18, the mean number of red blood cells [RBC, 10.5%] and platelets number [15%] were decreased. There was a significant relationship in RBC and platelets count. The mean diameter of RBC and nucleated red blood [NRBC] were compared in two groups. There was no significant relationship between experimental and control groups, except in NRBC diameter. Thus, the present data shows that ATRA may have negative effects on proliferation, differentiation and maturation of erythroid cells and platelets, without having any deleterious effects on the dimenation of RBC


Subject(s)
Animals, Laboratory , Tretinoin/pharmacology , Blood Cells/drug effects , Rats, Wistar , Embryonic Structures
17.
Article in English | WPRIM | ID: wpr-634604

ABSTRACT

Mouse blastocysts were exposed to doses of 0, 1 and 10 mumol/L retinoic acid (RA) for 24 h and the cytotoxic effect of RA on the mouse blastocysts in vitro was observed. FITC-labeled terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL-FITC) assay was employed to stain apoptotic cells and immunohistochemical S-P staining method was used to detect the expression of Fas protein in mouse blastocysts in vitro. The results showed that RA could induce apoptosis and increase the expression of Fas proteins of trophectoderm (TE) and inner cell mass (ICM) cells in blastocysts. Compared with the findings for the control blastocysts, exposure to RA (10 mumol/L) resulted in a more significant apoptosis and higher expression level of Fas proteins (P<0.01). It was concluded that RA could induce apoptosis, which may result in a significant reduction in the average number of total cells and the trophectoderm/inner cell mass in blastocysts and an increased expression of Fas protein, suggesting that RA had a cytotoxic effect on the growth and development of early embryos in mice.


Subject(s)
fas Receptor/biosynthesis , Apoptosis/drug effects , Blastocyst/cytology , Blastocyst/metabolism , Cell Culture Techniques/methods , Cells, Cultured , Gene Expression Regulation/drug effects , In Situ Nick-End Labeling , RNA, Messenger/metabolism , Tretinoin/pharmacology
18.
Article in English | WPRIM | ID: wpr-90620

ABSTRACT

9-cis-retinoic acid (9CRA) plays an important role in the immune response; this includes cytokine production and cell migration. We have previously demonstrated that 9CRA increases expression of chemokine receptors CCR1 and CCR2 in human monocytes. To better understand how 9CRA induces CCR1 and CCR2 expression, we examined the contribution of signaling proteins in human monocytic THP-1 cells. The mRNA and surface protein up-regulation of CCR1 and CCR2 in 9CRA-stimulated cells were weakly blocked by the pretreatment of SB202190, a p38 MAPK inhibitor, and PD98059, an upstream ERK inhibitor. Activation of p38 MAPK and ERK1/2 was induced in both a time and dose-dependent manner after 9CRA stimulation. Both p38 MAPK and ERK1/2 phosphorylation peaked at 2 h after a 100 nM 9CRA treatment. 9CRA increased calcium influx and chemotactic activity in response to CCR1-dependent chemokines, Lkn-1/CCL15, MIP-1alpha/CCL3, and RANTES/CCL5, and the CCR2-specific chemokine, MCP-1/CCL2. Both SB202190 and PD98059 pretreatment diminished the increased calcium mobilization and chemotactic ability due to 9CRA. SB202190 inhibited the expression and functional activities of CCR1 and CCR2 more effectively than did PD98059. Therefore, our results demonstrate that 9CRA transduces the signal through p38 MAPK and ERK1/2 for CCR1 and CCR2 up-regulation, and may regulate the pro-inflammatory process through the p38 MAPK and ERK-dependent signaling pathways.


Subject(s)
Calcium Signaling/drug effects , Cell Line , Chemokines/pharmacology , Chemotaxis, Leukocyte/drug effects , Enzyme Activation/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Flavonoids/pharmacology , Gene Expression Regulation/drug effects , Humans , Imidazoles/pharmacology , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Monocytes/drug effects , Pyridines/pharmacology , RNA, Messenger/genetics , Receptors, CCR1 , Receptors, CCR2 , Receptors, Chemokine/genetics , Tretinoin/pharmacology , p38 Mitogen-Activated Protein Kinases/metabolism
19.
Article in English | WPRIM | ID: wpr-174055

ABSTRACT

Skeletal muscle contains several precursor cells that generate muscle, bone, cartilage and blood cells. Although there are reports that skeletal muscle-derived cells can trans-differentiate into neural-lineage cells, methods for isolating precursor cells, and procedures for successful neural induction have not been fully established. Here, we show that the preplate cell isolation method, which separates cells based on their adhesion characteristics, permits separation of cells possessing neural precursor characteristics from other cells of skeletal muscle tissues. We term these isolated cells skeletal muscle-derived neural precursor cells (SMNPs). The isolated SMNPs constitutively expressed neural stem cell markers. In addition, we describe effective neural induction materials permitting the neuron-like cell differentiation of SMNPs. Treatment with retinoic acid or forskolin facilitated morphological changes in SMNPs; they differentiated into neuron-like cells that possessed specific neuronal markers. These results suggest that the preplate isolation method, and treatment with retinoic acid or forskolin, may provide vital assistance in the use of SMNPs in cell-based therapy of neuronal disease.


Subject(s)
Animals , Antigens, Differentiation/metabolism , Cell Adhesion , Cell Differentiation , Cell Lineage , Cell Separation , Cells, Cultured , Colforsin/pharmacology , Mice , Mice, Inbred ICR , Muscle, Skeletal/cytology , Neurons/cytology , Stem Cells/cytology , Tretinoin/pharmacology
20.
Article in English | WPRIM | ID: wpr-201424

ABSTRACT

Neurogenesis can be induced by pathological conditions such as cerebral ischemia. However the molecular mechanisms or modulating reagents of the reactive neurogenesis after the cerebral ischemia are poorly characterized. Retinoic acid (RA) has been shown to increase neurogenesis by enhancing the proliferation and neuronal differentiation of forebrain neuroblasts. Here, we examined whether RA can modulate the reactive neurogenesis after the cerebral ischemia. In contrast to our expectation, RA treatment decreased the reactive neurogenesis in subventricular zone (SVZ), subgranular zone (SGZ) and penumbral region. Furthermore, RA treatment also decreased the angiogenesis and gliosis in penumbral region.


Subject(s)
Animals , Brain/blood supply , Cell Differentiation , Cell Proliferation , Ischemic Attack, Transient/metabolism , Male , Neovascularization, Pathologic , Neuroglia/pathology , Neurons/pathology , Rats , Rats, Sprague-Dawley , Tretinoin/pharmacology
SELECTION OF CITATIONS
SEARCH DETAIL