RESUMO
Inhibition of matrix metalloproteinases (MMPs), which degrade collagen and elastin in the dermis of normal skin, is a key strategy for anti-skin aging. In this study, we identified five low-molecular-weight (LMW, <1 kDa) MMP-1-suppressive peptides in feather keratin hydrolysate (FKH) obtained by anaerobic digestion with an extremophilic bacterium. FKH was first subjected to ultrafiltration, followed by size-exclusion chromatography and liquid chromatography/electrospray ionization tandem mass spectrometry analysis. Chemically synthesized peptides identical to the sequences identified suppressed MMP expression in human dermal fibroblasts (HDFs). To investigate the impact of the MMP-1-suppressive peptides on the signaling pathway, we performed antibody array phosphorylation profiling of HDFs. The results suggested that the peptide GGFDL regulates ultraviolet-B-induced MMP-1 expression by inhibiting mitogen-activated protein kinases and nuclear factor κB signaling pathways as well as histone modification. Thus, LMW feather keratin peptides could serve as novel bioactive compounds to protect the skin against intrinsic and extrinsic factors.
Assuntos
Proteínas Aviárias/química , Plumas/química , Queratinas/química , Inibidores de Metaloproteinases de Matriz/química , Peptídeos/química , Envelhecimento da Pele/efeitos dos fármacos , Animais , Galinhas , Regulação para Baixo/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/efeitos da radiação , Humanos , Metaloproteinase 1 da Matriz/química , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/isolamento & purificação , Mapeamento de Peptídeos , Peptídeos/isolamento & purificação , Fosforilação , Hidrolisados de Proteína/química , Pele/efeitos dos fármacos , Pele/enzimologia , Pele/efeitos da radiação , Envelhecimento da Pele/genética , Envelhecimento da Pele/efeitos da radiação , Raios UltravioletaRESUMO
BACKGROUND/PURPOSE: The purpose of this study is to investigate the optimum combination of polyols and oils in moisturizing cosmetic products to improve the human skin moisturization and skin surface roughness. Polyols and oils are essential ingredients in skin care products, but it is still not understood how their concentrations affect their efficacy and sensory properties on human skin. We investigated the effect of polyol and oil concentration on skin properties by noninvasive methods. METHODS: The polyols consisted of glycerin and butylenes glycol in a ratio of 1:1 and the oils consisted of equal parts of hydrogenated polydecene, cethyl ethylhexanoate and pentaerythrityl tetraethylhexanoate. All cosmetic products were made in O/W emulsions in concentrations ranging from 0% to 30% for polyols and from 0% to 35% for oils. We investigated the effect on water content and skin surface roughness on the forearm after application of the cosmetic products. The skin water contents were measured by a Corneometer CM825 and the skin surface roughness by visual coring of skin surface biopsies in the scanning electron micrographs. RESULTS: In the first study, we found that the water content of the skin correlated highly with the polyol (up to 30%) and oil (up to 12%) concentrations, respectively. At two hours after application, the correlation coefficients were 0.971 and 0.985, respectively (P<0.01). Skin surface roughness not only showed a strong concentration dependence on polyols and oils (up to 6%). In the second study, we investigated the optimum combination of polyols and oils to improve the skin moisturization and skin surface roughness by the Response surface methodology. The water content of the skin surface was high in the ratio of polyol to oil (30:12 and 25:30). The skin surface roughness was improved considerably in the ratio of polyols to oil (30:6 and 30:35). CONCLUSIONS: Our results indicated that the skin surface properties were improved in the different ratios of their concentrations because they are influenced by not one ingredient but the interaction between polyols and oils. In this study, we could recommend the optimum concentration of polyols and oils to improve the skin surface properties. Further studies will be performed with other ingredients such as surfactants, lipids and so on.
Assuntos
Cosméticos/administração & dosagem , Óleos/administração & dosagem , Polímeros/administração & dosagem , Pele/efeitos dos fármacos , Pele/patologia , Adulto , Biópsia , Dermoscopia , Emolientes/administração & dosagem , Feminino , Humanos , Modelos Lineares , Masculino , Pele/metabolismo , Água/metabolismoRESUMO
Peroxisome proliferators activated receptors (PPARs) are a family of nuclear hormone receptors that heterodimer with the retinoid X receptor and function as transcriptional regulators of genes. Topically Applied PPAR-alpha agonists possess receptor mediated, pro-differentiating/anti-proliferative effects, lipid metabolism stimulation, and anti-inflammatory activity, which suggest that they could be beneficial for the treatment of a variety of cutaneous diseases. Hyaluronan (HA), a high-molecular-weight linear glycosaminoglycan consisting of alternating D: -glucuronic acid and N-acetyl-D: -glucosamine residues, is one of the major extracellular matrix components in skin. Among the family of HA synthase genes (HAS1, 2, 3) so far identified, one group has demonstrated that the expressions of HAS2 and HAS3 play crucial roles in the regulation of HA synthesis in human skin fibroblasts and keratinocytes, respectively, but the precise regulatory mechanisms are still unknown. We examine Musk T called Ethylene brassylate, Astratone or 1,4-Dioxacycloheptadecane-5,17-dione, which used as just a perfume ingredient, plays a role as PPAR-alpha ligand in vitro and stimulates skin barrier recovery, ceramide synthesis, beta-Glucocerebrosidase, involucrin expression in epidermis in vivo; and examine that Musk T stimulates HAS expression and HA synthesis in human skin fibroblast. Through these experiments, we conclude that Musk T is PPAR-alpha ligand, effects on keratinocyte differentiation, intercellular lipid synthesis in epidermis, HA synthesis stimulation in dermis.
Assuntos
Epiderme/metabolismo , Éteres Cíclicos/farmacologia , Ácidos Graxos Monoinsaturados/farmacologia , Homeostase/fisiologia , Ácido Hialurônico/metabolismo , PPAR alfa/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Chlorocebus aethiops , Epiderme/efeitos dos fármacos , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Proteínas Filagrinas , Glucuronosiltransferase/metabolismo , Homeostase/efeitos dos fármacos , Humanos , Hialuronan Sintases , Proteínas de Filamentos Intermediários/metabolismo , Queratinócitos/citologia , Queratinócitos/metabolismo , Camundongos , Camundongos Pelados , Precursores de Proteínas/metabolismo , RNA Mensageiro/metabolismoRESUMO
Naturally occurring triterpenoids such as oleanolic acid (OA) and ursolic acid (UA) are known to have anti-inflammatory and anticarcinogenic activities in some types of cells. Although it has been reported that UA increases the amount of ceramide in keratinocytes, there is little study on the mechanism of triterpenoids involved in the differentiation of keratinocytes as well as their effects on epidermal permeability barrier. A study was therefore conducted to determine whether OA and UA could stimulate the differentiation of epidermal keratinocytes through peroxisome proliferator-activated receptor (PPAR)-alpha activation. This work was then extended to investigate the rate of formation of cornified envelope as a marker in the terminal differentiation of keratinocytes and the amount of transglutaminase in human keratinocytes treated with OA and UA. It was shown that OA induced the differentiation of keratinocytes, whereas UA had little effect. In addition, reporter gene assay using PPAR response element activity demonstrated that OA might be related to the increase of PPAR-alpha activity in CV-1 cells. Moreover, it enhanced the recovery of epidermal permeability barrier function as well as increased ceramides in epidermis after topical application. We therefore propose that the effect of OA on the stimulation of differentiation in epidermal keratinocytes seems to be highly related to activation of PPAR-alpha.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Queratinócitos/efeitos dos fármacos , Ácido Oleanólico/farmacologia , PPAR alfa/efeitos dos fármacos , Triterpenos/farmacologia , Animais , Diferenciação Celular/fisiologia , Linhagem Celular , Linhagem Celular Transformada , Ceramidas/metabolismo , Epiderme/metabolismo , Humanos , Queratinócitos/fisiologia , Camundongos , PPAR alfa/fisiologia , Permeabilidade/efeitos dos fármacos , Plantas/química , Ácido UrsólicoRESUMO
A native-feather-degrading thermophilic anaerobe was isolated from a geothermal hot stream in Indonesia. Isolate AW-1, identified as a member of the species Fervidobacterium islandicum, was shown to degrade native feathers (0.8%, w/v) completely at 70 degrees C and pH 7 with a maximum specific growth rate (0.14 h(-1)) in Thermotoga- Fervidobacterium(TF) medium. After 24 h of culture, feather degradation led to an increase in free amino acids such as histidine, cysteine and lysine. Moreover, nutritionally essential amino acids such as tryptophan and methionine, which are rare in feather keratin, were also produced as microbial metabolites. A homomultimeric membrane-bound keratinolytic protease (>200 kDa; 97 kDa subunits) was purified from a cell extract of F. islandicum AW-1. The enzyme exhibited activity toward casein and soluble keratin optimally at 100 degrees C and pH 9, and had a half-life of 90 min at 100 degrees C. The enzyme showed higher specific activity for the keratinous substrates than other proteases and catalyzed the cleavage of peptide bonds more rapidly following the reduction of disulfide bridges in feather keratin by 10 mM dithiothreitol. Therefore, the enzyme from F. islandicum AW-1 is a novel, thermostable keratinolytic serine protease.