RESUMEN
BACKGROUND: Hydration is an important factor to promote skin barrier function, metabolism, and appearance. In this process, the presence of aquaglyceroporins, envelope and lipid synthesis, and metabolism proteins are essential to provide greater corneocyte cohesion and to form a barrier avoiding transepidermal water loss. OBJECTIVE: We evaluated the effects of a new topical pigment-free agent containing an Anadenanthera colubrina polysaccharide-rich dermocosmetic preparation (ACP) on the aquaporin-3 (AQP-3), filaggrin (FLG), involucrin (INV), glucocerebrosidase (GBA), and elongation of very-long-chain fatty acid (ELOVL) proteins production in skin human fragments, as well as on the transepidermal water loss in a double-blind placebo-controlled clinical trial. METHODS: AQP3, FLG, INV, GBA, and ELOVL3 levels were measured by immunofluorescence analysis in human skin explants. Clinical trial was conducted to evaluate the effects of ACP 1% and ACP 3% on the transepidermal water loss (TEWL). RESULTS: Image and statistical analysis showed that ACP 3% significantly increased at 90% the expression of AQP3. Similarly, ACP 3% was able to promote a significant increase of 68% and 51% in FLG and INV, respectively. ACP 3% produced no effects on the GBA and ELOVL3 proteins. Transepidermal water loss was significantly reduced in human volunteers under treatment with ACP 1% and ACP 3%. CONCLUSION: ACP reduced transepidermal water loss in a clinical trial, promoting human skin hydration. These effects were related to modulation of the AQP3, FLG, and INV as evidenced by immunofluorescence assay. This way, A colubrina polysaccharide-rich phytopharmaceutical preparation is an effective additive product to skin hydration.
Asunto(s)
Colubrina , Proteínas Filagrina , Humanos , Preparaciones de Plantas , Polisacáridos/metabolismo , Piel/metabolismo , Agua/metabolismo , Pérdida Insensible de AguaRESUMEN
SummaryThe objectives were to develop an effective protocol for transfection of ovine secondary follicles and to assess the effect of attenuating aquaporin 3 (AQP3) using a small interfering RNA (siRNA-AQP3) on antrum formation and follicular growth in vitro. Various combinations of Lipofectamine® volumes (0.5, 0.75 or 1.0 µl), fluorescent oligonucleotide (BLOCK-iT ™) concentrations (3.18, 27.12 or 36.16 nM) and exposure times (12, 14, 16, 18 or 20 h) were tested. The BLOCK-iT™ was replaced by siRNA-AQP3 in the transfection complex. Ovine secondary follicles were isolated and cultured in vitro for 6 days using standard protocols. Follicles were transfected on day 0 or 3 or on both days (0 and 3) and then cultured for an additional 3 or 6 days. As revealed by the fluorescence signal, the Lipofectamine®/BLOCK-iT™ complex (0.75 µl + 27.12 nM by 12 h of incubation) crossed the basement membrane and granulosa cell and reached the oocytes. In general, the rate of intact follicles was higher and the rate of antrum formation was lower in transfected follicles compared with control follicles. In conclusion, ovine secondary follicles can be successfully transfected during in vitro culture, and siRNA-mediated attenuation of AQP3 gene reduced antrum formation of secondary follicles.
Asunto(s)
Acuaporina 3/genética , Folículo Ovárico/fisiología , Transfección/métodos , Animales , Acuaporina 3/metabolismo , Técnicas de Cultivo de Célula , Femenino , Técnicas de Silenciamiento del Gen , Lípidos , Folículo Ovárico/crecimiento & desarrollo , Interferencia de ARN , OvinosRESUMEN
Several aquaporins (AQPs) are expressed in extravillous (EVT) and villous trophoblast cells. Among them, AQP3 is the most abundant AQP expressed in chorionic villi samples from first trimester, followed by AQP1 and AQP9. Although AQP3 expression persists in term placentas, it is significantly decreased in placentas from preeclamptic pregnancies. AQP3 is involved in the migration of different cell types, however its role in human placenta is still unknown. Here, we evaluated the role of AQP3 in the migration of EVT cells during early gestation. Our results showed that Swan 71â¯cells expressed AQP1, AQP3 and AQP9 but only the blocking of AQP3 by CuSO4 or the silencing of its expression by siRNA significantly attenuates EVT cell migration. Our work provides evidence that AQP3 is required for EVT cell migration and suggests that an altered expression of placental AQP3 may produce failures in placentation such as in preeclampsia.
Asunto(s)
Acuaporina 3/antagonistas & inhibidores , Movimiento Celular , Trofoblastos/citología , Trofoblastos/metabolismo , Acuaporina 1/antagonistas & inhibidores , Acuaporina 1/metabolismo , Acuaporina 3/genética , Acuaporina 3/metabolismo , Línea Celular , Movimiento Celular/genética , Silenciador del Gen , Humanos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Cicatrización de HeridasRESUMEN
The mRNA expression and localization of Aquaporin 3 (AQP3) were investigated in the ovarian follicles of ewes at different stages of development (primordial, primary, secondary, small, and large antral). The gene expression was quantified by qPCR, while the protein identification and localization were determined by Western blot and immunohistochemistry, respectively. Analysis revealed that AQP3 mRNA was detected only in the antral follicles, whereas the protein expression was detected in the oocyte and granulosa cells in all stages of follicular development. The latter observation suggests that the presence of AQP3 in follicles of all categories, especially in the antral follicles, provides novel insights on the mechanisms that regulate the flow of water between cells during the formation of antral follicles in sheep.