RESUMEN
Abnormal melanin synthesis results in several hyperpigmentary disorders such as freckles, melanoderma, age spots, and other related conditions. In this study, we investigated the antimelanogenic effects of an extract from the microalgae Chlamydomonas reinhardtii (CE) and potential mechanisms responsible for its inhibitory effect in B16F10, normal human epidermal melanocyte cells, and human skin-equivalent models. The CE extract showed significant dose-dependent inhibitory effects on α-melanocyte-stimulating, hormone-induced melanin synthesis in cells. Additionally, the CE extract exhibited suppressive effects on the mRNA and protein expression of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2. The CE extract also inhibited the phosphorylation of protein kinase A and extracellular signal-related kinase, which function as upstream regulators of melanogenesis. Using a three-dimensional, reconstructed pigmented epidermis model, the CE-mediated, anti-pigmentation effects were confirmed by Fontana-Masson staining and melanin content assays. Taken together, CE extract can be used as an anti-pigmentation agent.
Asunto(s)
Chlamydomonas reinhardtii/química , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Pigmentación/efectos de los fármacos , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Línea Celular Tumoral/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Descubrimiento de Drogas , Células Epidérmicas/efectos de los fármacos , Células Epidérmicas/patología , Epidermis/efectos de los fármacos , Epidermis/patología , Quinasas MAP Reguladas por Señal Extracelular , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Péptidos y Proteínas de Señalización Intracelular/farmacología , Oxidorreductasas Intramoleculares/metabolismo , Melaninas/metabolismo , Melanoma , Glicoproteínas de Membrana/metabolismo , Ratones , Factor de Transcripción Asociado a Microftalmía/metabolismo , Monofenol Monooxigenasa/metabolismo , Oxidorreductasas/metabolismo , Fosforilación , ARN Mensajero/efectos de los fármacos , Piel , alfa-MSH/metabolismoRESUMEN
Arctiin, a lignin isolated from Arctium lappa, exhibits a variety of biological effects, including antiviral, antiinflammatory, and antiproliferative actions, in mammalian cells. In a previous study, arctiin was demonstrated to induce procollagen type I synthesis and exhibited protective effects against ultraviolet B (UVB) radiation in normal human dermal fibroblasts (nHDFs). However, the underlying molecular mechanism of arctiinmediated collagen synthesis remains unknown. In the present study, the mechanism for increased expression of collagen type 1α 1 chain (COL1A1) mRNA in arctiininduced nHDFs was identified. The expression of microRNA378b (miR378b), downregulated by arctiin, was correlated with the expression of sirtuin6 (SIRT6) mRNA, a regulator of COL1A1 mRNA. Furthermore, it was revealed that arctiin protected the UVB radiationmediated decrease in COL1A1 mRNA expression, through the miR378b/SIRT6 signaling pathway. In conclusion, these results suggest that arctiin regulates COL1A1 through the miR378bSIRT6 axis.
Asunto(s)
Colágeno Tipo I/metabolismo , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Furanos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Glucósidos/farmacología , MicroARNs/genética , ARN Mensajero/genética , Sirtuinas/genética , Supervivencia Celular , Células Cultivadas , Cadena alfa 1 del Colágeno Tipo I , Dermis/citología , Regulación de la Expresión Génica/efectos de la radiación , Humanos , Extractos Vegetales/farmacología , Interferencia de ARN , Rayos UltravioletaRESUMEN
Dermal papilla (DP) is a pivotal part of hair follicle, and the smaller size of the DP is related with the hair loss. In this study, we investigated the effect of titrated extract of Centella asiatica (TECA) on hair growth inductive property on 3D spheroid cultured human DP cells (HDP cells). Significantly increased effect of TECA on cell viability was only shown in 3D sphered HPD cells, not in 2D cultured HDP cells. Also, TECA treatment increased the sphere size of HDP cells. The luciferase activity of STAT reporter genes and the expression of STAT-targeted genes, SOCS1 and SOCS3, were significantly decreased. Also, TECA treatment increased the expression of the hair growth-related signature genes in 3D sphered HDP cells. Furthermore, TECA led to downregulation of the level of phosphorylated STAT proteins in 3D sphered HDP cells. Overall, TECA activates the potential of hair inductive capacity in HDP cells.
Asunto(s)
Centella/química , Dermis/citología , Cabello/efectos de los fármacos , Cabello/crecimiento & desarrollo , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Esferoides Celulares/citología , Esferoides Celulares/metabolismoRESUMEN
Ultraviolet (UV) light exposure causes skin photoaging, which is known to be preventable and controllable by application of UV-protective agents. In this study, we demonstrated, for the first time, that the extract of microalgae Arthrospira platensis has a reverse effect on UV-induced photodamage such as loss of cell viability, cellular senescence, DNA damage, and collagen destruction in dermal fibroblasts. Forty-eight extracts were prepared from the cell biomass by controlling culture light conditions, extract solvents, and disruption methods. Then, we analyzed their cytotoxicities using WST-1 assay and separated low and high cytotoxic extracts with normal human dermal fibroblasts (nHDFs). Using the low cytotoxic extracts, we performed UVB protection assay and selected the most effective extract demonstrating protective effect against UVB-induced nHDF damage. Flow cytometric analysis and senescence-associated (SA) ß-galactosidase assay showed that pretreatment with the extract reversed UVB-induced G2/M phase cell cycle arrest and senescence in nHDFs. Furthermore, UVB-induced DNA damage in nHDFs, such as cyclobutane pyrimidine dimer formation, was significantly suppressed by the extract. Further, quantitative real-time PCR experiments revealed that the extract significantly inhibited UVB-induced upregulation of matrix metalloproteinase 1 (MMP1) and MMP3 expression in nHDFs. Therefore, we concluded that the microalgae extract can be a potential anti-photoaging agent.
Asunto(s)
Senescencia Celular/efectos de la radiación , Daño del ADN/efectos de los fármacos , Metaloproteinasa 1 de la Matriz/metabolismo , Extractos Vegetales/química , Sustancias Protectoras/farmacología , Spirulina/química , Rayos Ultravioleta , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Senescencia Celular/efectos de los fármacos , Daño del ADN/efectos de la radiación , Dermis/citología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de la radiación , Humanos , Puntos de Control de la Fase M del Ciclo Celular/efectos de los fármacos , Puntos de Control de la Fase M del Ciclo Celular/efectos de la radiación , Metaloproteinasa 1 de la Matriz/genética , Extractos Vegetales/farmacología , Sustancias Protectoras/química , Dímeros de Pirimidina/efectos de la radiación , Spirulina/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Melanogenesis is the process of generating pigmentation via melanin synthesis and delivery. Three key enzymes, tyrosinase, tyrosinase-related protein 1 (TRP1) and TRP2, metabolize melanin from L-tyrosine. Melanin synthesizing enzymes are regulated by microphthalmia-associated transcription factor (MITF). The titrated extract of Centella asiatica (TECA) contains the major components asiatic acid, asiaticoside and madecassic acid. The present study revealed that TECA reduces the melanin content in melanocytes. Moreover, the asiaticoside contained in TECA modulated melanogenesis by inhibiting tyrosinase mRNA expression. The decrease in tyrosinase mRNA levels was mediated through MITF. Uniquely, asiaticoside inhibited MITF by decreasing its DNA binding affinity. In conclusion, the results of the present study indicate that asiaticoside treatment may have beneficial effects in hyperpigmentation diseases or for skin whitening.
Asunto(s)
Antiinfecciosos/farmacología , Centella/química , Melaninas/biosíntesis , Melanocitos/efectos de los fármacos , Triterpenos/farmacología , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Centella/metabolismo , ADN/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Melanocitos/metabolismo , Melanoma Experimental/metabolismo , Melanoma Experimental/patología , Ratones , Factor de Transcripción Asociado a Microftalmía/metabolismo , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/metabolismo , Unión ProteicaRESUMEN
Ultraviolet (UV) radiation induces severe alterations in the molecular and cellular components of normal human dermal fibroblast (NHDF) cells by disrupting many intracellular transduction cascades. Although UV responses have been well documented at the genome and proteome levels, UV protective effects have not been elucidated at these levels. The aim of the present study was to demonstrate that arctiin, a phytochemical isolated from the plant Arctium lappa, induced a protective effect against UVB radiation by changing microRNA (miRNA) expression profiles. Using flow cytometry, and water-soluble tetrazolium salt (WST-1)-based cell viability, wound healing, and DNA repair assays we showed that pretreatment with arctiin prior to UVB irradiation reduced UVB-induced apoptosis, cell migration defects, and DNA damage in NHDF cells. It was also found that arctiininduced UVB protection is associated with altered miRNA expression profiles. Bioinformatic analysis revealed that the deregulated miRNAs were functionally involved in mitogen-activated protein kinase (MAPK) signaling and cancer signaling pathways. The results suggest that arctiin acts as a UVB protective agent by altering specific miRNA expression in NHDF cells.
Asunto(s)
Fibroblastos/efectos de los fármacos , Furanos/administración & dosificación , Glucósidos/administración & dosificación , Protectores contra Radiación/administración & dosificación , Transducción de Señal/efectos de los fármacos , Arctium/química , Fibroblastos/efectos de la radiación , Furanos/química , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/efectos de la radiación , Glucósidos/química , Humanos , MicroARNs/metabolismo , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Protectores contra Radiación/química , Transducción de Señal/efectos de la radiación , Piel/citología , Piel/efectos de los fármacos , Piel/efectos de la radiación , Rayos UltravioletaRESUMEN
microRNAs (miRNAs) have been shown to function as primary regulators of a variety of biological processes, including proliferation, differentiation and apoptosis in human keratinocytes. However, the biological significance of miRNAs in the defense against oxidative stress in keratinocytes remains to be elucidated. In this study, we demonstrate that oridonin, a diterpenoid compound isolated from Rabdosia rubescens with established antioxidant properties, protects HaCaT human keratinocytes from oxidative stress induced by exposure to hydrogen peroxide (H2O2). Our data demonstrate that low doses of oridonin (1-5 µM) protect keratinocytes against H2O2-induced apoptosis in a concentration- and time-dependent manner. Moreover, as shown by our results, oridonin markedly decreased H2O2-induced reactive oxygen species production in HaCaT cells. Oridonin mediated these effects by altering miRNA expression. Bioinformatics analysis identified several putative target genes of the differentially expressed miRNAs. Assessment of their gene ontology annotation revealed that these target genes are likely involved in cell growth and inhibition of apoptosis. Thus, the data from this study establish a role for miRNAs in mediating oridonin-induced protective effects against oxidative stress in human keratinocytes.
Asunto(s)
Diterpenos de Tipo Kaurano/farmacología , Peróxido de Hidrógeno/efectos adversos , Queratinocitos/efectos de los fármacos , MicroARNs/metabolismo , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Biología Computacional , Humanos , Queratinocitos/metabolismo , MicroARNs/genética , Análisis por Micromatrices , Sustancias Protectoras/farmacología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
This study aimed to evaluate the protective effects of Centella asiatica (C. asiatica) against ultraviolet B (UVB) damage in human keratinocytes using microRNA (miRNA) expression profiling analysis. Titrated extract of C. asiatica (TECA) demonstrated low cytotoxicity in normal human HaCaT keratinocytes only at low doses (<5 µg/ml). UVB (50 mJ/cm2) irradiation significantly decreased cell viability, and TECA treatment decreased the UVB toxicity. By using miRNA microarrays, we determined that 72 miRNAs had an altered expression following TECA treatment in UVB-irradiated keratinocytes (46 upregulated and 26 downregulated). Using an miRNA target gene prediction tool and Gene Ontology (GO) analysis, we determined that miRNAs with altered expression were functionally related with the inhibition of apoptosis and cell proliferation. Overall, these results provide meaningful information to facilitate the understanding of TECA-mediated UVB protection in human keratinocytes.
Asunto(s)
Centella/química , MicroARNs/genética , Extractos Vegetales/farmacología , Tolerancia a Radiación/efectos de los fármacos , Protectores contra Radiación/farmacología , Transcriptoma/efectos de los fármacos , Apoptosis/genética , Línea Celular , Proliferación Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Expresión Génica/efectos de los fármacos , Expresión Génica/efectos de la radiación , Humanos , Queratinocitos , MicroARNs/metabolismo , Anotación de Secuencia Molecular , Análisis de Secuencia por Matrices de Oligonucleótidos , Transcriptoma/efectos de la radiación , Rayos UltravioletaRESUMEN
The titrated extract of Centella asiatica (TECA) is a reconstituted mixture comprising of asiatic acid, madecassic acid, asiaticoside and madecassoside, and is used as a therapeutic agent in wound healing and also as an anti-microbial, anticancer and anti-aging agent. Although these properties and the associated cell signaling pathways have been elucidated, the cellular mechanism of anti-photoaging upon ultraviolet (UV) exposure in normal human dermal fibroblasts (NHDFs) remains unknown. In this study, we investigated the photoprotective role of TECA via microRNA (miRNA) expression profiling analysis. Low dose of TECA did not exhibit toxicity and showed a protective effect against UVB irradiation in NDHFs. miRNA microarray experiments revealed that specific miRNAs were altered by TECA stimulation in UVB-irradiated NHDFs. Functional bioinformatic analysis showed that the putative target genes of the altered miRNAs were associated with the positive regulation of cell proliferation, anti-apoptosis, small GTPase- and Ras-mediated signal transduction and activation of MAPKK. Therefore, these results suggest that TECA may serve as a potential natural chemoprotective agent against UVB-mediated damage in NHDFs through changes in the expression of specific miRNAs.
Asunto(s)
Fibroblastos/efectos de los fármacos , MicroARNs/genética , Extractos Vegetales/farmacología , Protectores contra Radiación/farmacología , Transcriptoma/efectos de los fármacos , Triterpenos/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Centella , Dermis/citología , Fibroblastos/efectos de la radiación , Humanos , MicroARNs/metabolismo , Interferencia de ARN , Rayos UltravioletaRESUMEN
Centella asiatica (C. asiatica) is a pharmacological plant in South Asia. It has been demonstrated that C. asiatica extracts containing various pentacyclic triterpenes exert healing effects, especially wound healing and collagen synthesis in skin. However, there are few studies on the effect of C. asiatica extracts on stress-induced premature senescence (SIPS). To determine whether H(2) O(2) -induced senescence is affected by C. asiatica extracts, we performed senescence analysis on cultured human dermal fibroblasts (HDFs). We also analysed whole gene expression level using microarrays and showed that 39 mRNAs are differentially expressed in H(2) O(2) -induced HDFs with and without treatment with C. asiatica extracts. These genes regulate apoptosis, gene silencing, cell growth, transcription, senescence, DNA replication and the spindle checkpoint. Differential expression of FOXM1, E2F2, MCM2, GDF15 and BHLHB2 was confirmed using semi-quantitative PCR. In addition, C. asiatica extracts rescued the H(2) O(2) -induced repression of replication in HDFs. Therefore, the findings presented here suggest that C. asiatica extracts might regulate SIPS by preventing repression of DNA replication and mitosis-related gene expression.
Asunto(s)
Senescencia Celular/efectos de los fármacos , Dermis/citología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Triterpenos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Proteínas de Ciclo Celular/genética , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Centella , Inhibidor p16 de la Quinasa Dependiente de Ciclina , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Replicación del ADN/efectos de los fármacos , Regulación hacia Abajo/genética , Factor de Transcripción E2F2/genética , Fibroblastos/metabolismo , Proteína Forkhead Box M1 , Factores de Transcripción Forkhead/genética , Expresión Génica/efectos de los fármacos , Expresión Génica/genética , Perfilación de la Expresión Génica , Factor 15 de Diferenciación de Crecimiento/genética , Proteínas de Homeodominio/genética , Humanos , Componente 2 del Complejo de Mantenimiento de Minicromosoma , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Extractos Vegetales , Reacción en Cadena de la Polimerasa , Proteína p53 Supresora de Tumor/metabolismo , Regulación hacia Arriba/genéticaRESUMEN
Iris nertschinskia, an ornamental plant, is utilized in traditional East Asian medicine for the treatment of skin diseases. However, the biological activity underlying its therapeutic effects remains to be established. In this study, we investigated the anti-tumor effect of the plant extract on MCF7 human breast cancer cells. An ethanol extract of Iris nertschinskia triggered cell death in a dose-dependent manner. Moreover, treatment with the extract promoted p53 phosphorylation in MCF7 cells. Increased phosphorylation of p53, in turn, led to induction of Bax protein, a key regulator of p53-dependent apoptotic cell death, as well as of caspase-7 cleavage in MCF7 cells. Consistently, cells treated with p53-specific siRNA or the caspase inhibitor, Z-VAD, resisted apoptotic cell death induced by the Iris nertschinskia extract. Our results suggest that p53 sensitizes tumor cells to the ethanol extract of Iris nertschinskia by Bax protein induction and caspase-dependent apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Neoplasias de la Mama/metabolismo , Género Iris/química , Extractos Vegetales/farmacología , Proteína p53 Supresora de Tumor/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Caspasa 7/metabolismo , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Etanol/química , Femenino , Humanos , Fosforilación/efectos de los fármacos , Extractos Vegetales/química , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Black rice (Oryza sativa L. var. japonica) has been used in folk medicine in Asia. To understand the effects of black rice hydrolyzed peptides (BRP) from germinated black rice, we assessed the expression levels of about 20,000 transcripts in BRP-treated HaCaT keratinocytes using human 1A oligo microarray analysis. As a result, the BRP treatment showed a differential expression ratio of more than 2-fold: 745 were activated and 1,011 were repressed. One of the most interesting findings was a 2-fold increase in hyaluronan synthase 2 (HAS2) gene expression by BRP. Semiquantitative RT-PCR showed that BRP increased HAS2 mRNA in dose-dependent manners. ELISA showed that BRP effectively increased hyaluronan (HA) production in HaCaT keratinocytes.