RESUMEN
Skin aging is induced through complex biological processes in human skin caused by proteolysis of collagen and elastin, two structural proteins of the dermal extracellular matrix (ECM). Collagen and elastin degradation can induce the expression of matrix metalloproteinases (MMPs), as well as that of a family of zinc-dependent endopeptidases that play critical roles in skin aging. Moreover, elastase is a metalloproteinase which acts on the degradation of elastin in skin aging, and is also involved in the inhibition or the repair of wrinkle formation. Extract of the mycelium of Tricholoma matsutake (T. matsutake), or pine mushroom, is widely distributed in Asian countries. The extract is from the natural biomaterial of the mushroom which is rich in polysaccharides, including ß-glucan. This extract has shown potent bioactive antioxidant, immunomodulatory and antitumoral properties. In the present study, we investigated whether the extract of the mycelium of T. matsutake has effects on elastase activity, as well as on the expression of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) and MMP-1 under basal conditions. Our results revealed that the extract of the mycelium of T. matsutake significantly decreased elastase activity in a dose-dependent manner and reduced the levels of MMP-1 and MMP-3. On the other hand, the expression of TIMP-1 and tropoelastin was increased in fibroblasts treated with the extract of the mycelium of T. matsutake. However, collagent expression was not affected. In addition, our results demonstrated that the extract of the mycelium of T. matsutake inhibited the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced MMP-1 expression and suppressed TPA-induced p38 activity. Therefore, the inhibitory effects of the extract of the mycelium of T. matsutake on MMP-1 induction are mediated by the inhibition of p38 in human fibroblasts. Our data suggest that the extract of the mycelium of T. matsutake may prove to be an effective biomaterial for anti-wrinkle treatment, as it can obstruct the degradation of the dermal ECM.
Asunto(s)
Productos Biológicos/farmacología , Fibroblastos/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Metaloproteinasa 1 de la Matriz/genética , Elastasa Pancreática/antagonistas & inhibidores , Acetato de Tetradecanoilforbol/farmacología , Adulto , Productos Biológicos/aislamiento & purificación , Western Blotting , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Células Cultivadas , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Relación Dosis-Respuesta a Droga , Fibroblastos/metabolismo , Humanos , Metaloproteinasa 1 de la Matriz/metabolismo , Micelio/química , Elastasa Pancreática/metabolismo , Fosforilación/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Piel/citología , Inhibidor Tisular de Metaloproteinasa-1/genética , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Tricholoma/química , Tropoelastina/clasificación , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Ablative fractional carbon dioxide (CO2) lasers have been widely used for several types of cosmetic dermatosis. A number of previous studies have evaluated this technique in animals or human beings by observing morphologic changes using an invasive modality such as skin biopsy. In this study, we assessed in vivo skin changes after CO2 ablative fractional laser treatment in a mouse model using noninvasive imaging modalities (Folliscope(®) and Visioscan 98(®)), and each results was compared with data from histologic examination. An ablative fractional CO2 laser was applied with different pulse energy between 7 to 35 mJ/microspot. As results of above methods, we also confirmed that the CO2 ablative fractional laser generated injuries with increasing width and depth with increasing pulse energy. Although numerous papers have described application of this laser in vivo skin specimens, our study evaluated the feasibility of using relative noninvasive imaging modalities for assessing the outcome of laser ablation. Based on our data, we suggest that these technologies may be useful alternative modalities for assessing laser ablation that are easier to perform and less invasive than skin biopsy.
Asunto(s)
Láseres de Gas , Animales , Femenino , Ratones , Ratones Endogámicos BALB C , Modelos AnimalesRESUMEN
Growth factors are important for regulating a variety of cellular processes and typically act as signaling molecules between cells. In the present study, we examined the mechanisms underlying the inhibitory effects of mixtures of recombinant growth factors (MRGFs) on nitric oxide (NO) and pro-inflammatory cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We also examined whether these effects are mediated through the mitogenactivated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signal transduction pathways. NO production was assessed by measuring nitrite acucmulation using the Greiss reaction. Cytokine concentrations were measured using respective ELISA kits for each cytokine. Our results revealed that the MRGFs significantly attenuated the LPS-induced production of pro-inflammatory cytokines and NO in a dose-dependent manner. To elucidate the mechanisms underlying the inhibitory effects of MRGFs, we examined the effects of the LPS-induced phosphorylation of MAPKs and the activation of the NF-κB signaling pathway on the stabilization of NF-κB nuclear translocation and inhibitory factor-κB (IκB) degradation. Western blot analysis was performed to determine the total and phosphorylated levels of ERK, as well as the nuclear translocation of NF-κB, and IκB phosphorylation and degradation. Our results demonstrated that treatment with MRGFs resulted in a reduction in the phosphorylation of the ERK and NF-κB signaling pathways, whereas the phosphorylation of JNK and p38 was not affected. Taken together, our results suggest that MRGFs inhibit the production of pro-inflammatory cytokines and NO by downregulating inducible NO synthase gene expression and blocking the phosphorylation of the ERK and NF-κB signaling pathways. These findings may provide direct evidence of the potential application of MRGFs in the prevention and treatment of inflammatory diseases.
Asunto(s)
Citocinas/biosíntesis , Factor de Crecimiento Epidérmico/deficiencia , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Factor I del Crecimiento Similar a la Insulina/deficiencia , Insulina/deficiencia , Óxido Nítrico/metabolismo , Animales , Citocinas/efectos de los fármacos , Factor de Crecimiento Epidérmico/administración & dosificación , Inflamación/inducido químicamente , Insulina/administración & dosificación , Factor I del Crecimiento Similar a la Insulina/administración & dosificación , Lipopolisacáridos/toxicidad , Errores Innatos del Metabolismo , Ratones , Proteínas Quinasas Activadas por Mitógenos/biosíntesis , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Transducción de Señal/efectos de los fármacosRESUMEN
The potential role of topical valproate (VPA) in hair regrowth has been recently suggested. However, safety reports of VPA as a topical formulation are lacking. Therefore, in the present study, we investigated whether VPA causes skin irritation in humans. We first performed a cell viability test and showed that VPA did not exhibit toxicity toward HaCaT keratinocytes, fibroblasts, and RBL-3H mast cells. We then performed clinical patch test and skin irritation test through transdermal drug delivery with the help of microneedle rollers. No significant findings were obtained in the clinical patch test. In the skin irritation test, only 1 patient showed erythema at 1 hr, but the irritation reaction faded away within a few hours. Erythema and edema were not observed at 24 hr. We concluded that VPA has minimal potential to elicit skin irritation. Therefore, we consider that VPA can safely be applied to human skin.