Espada J, Matabuena M, Salazar N, Lucena S, Kourani O, Carrasco E, Calvo M, Rodríguez C, Reyes E, González S, Juarranz A
Department of Biology, Faculty of Sciences, Universidad Autónoma de Madrid, Madrid, Spain.
Int J Cosmet Sci. 2015 Feb;37(1):41-55. doi: 10.1111/ics.12167. Epub 2014 Oct 27.
The search of substances that minimize cutaneous ageing has increased in the last few years. Previous studies have described the regenerative properties of the secretion of the mollusc Cryptomphalus aspersa (C. aspersa) when applied topically.
We evaluate the in vitro effects of a new product derived from the eggs of C. aspersa, IFC-CAF, on cell proliferation, migration, distribution of cytoskeletal proteins, production of extracellular components as well as its ability to prevent cutaneous ageing because of intrinsic or extrinsic factors (exposure to UVB) by determination of ageing markers.
We have used the human keratinocyte cell line (HaCaT cells), primary dermal fibroblasts (HDF) and senescent dermal fibroblasts (SHDF). The effects of the compound on cell proliferation and on the cell cycle were determined by the MTT colorimetric assay, estimation of total protein and/or trypan blue test and by flow cytometry, respectively. We also studied cell migration using the wound-healing migration assay, whereas ELISA assays, Western Blot and immunofluorescence microscopy were carried out to test the expression of proteins related to cytoskeleton, extracellular matrix and with ageing.
We have found that IFC-CAF does not promote proliferation but induces migration of HaCaT, HDF and SHDF in a time- and dose-dependent manner; a better organization of cytoskeletal proteins (F-actin and vimentin) and promotes the production of extracellular components (fibronectin, collagen 1 and MMPs) and the adhesion to cell-substrate vinculin protein. IFC-CAF also prevents cutaneous ageing. The treatment decreases the expression of the ageing-related markers b-Gal, p53 and p16INK4 in SDDF cells, and improves cell survival after UVB irradiation and nuclear repair in HaCaT cells.
IFC-CAF has regenerative properties and protects against ageing factors being, therefore, a potential therapeutic agent for treating or preventing skin ageing.
在过去几年中，寻找能使皮肤衰老最小化的物质的研究有所增加。先前的研究描述了将软体动物隐孔螺（C. aspersa）的分泌物局部应用时的再生特性。
我们评估了一种源自C. aspersa卵的新产品IFC-CAF对细胞增殖、迁移、细胞骨架蛋白分布、细胞外成分产生的体外作用，以及通过测定衰老标志物来评估其预防因内在或外在因素（暴露于UVB）导致皮肤衰老的能力。
我们使用了人角质形成细胞系（HaCaT细胞）、原代表皮成纤维细胞（HDF）和衰老的真皮成纤维细胞（SHDF）。分别通过MTT比色法、总蛋白估计和/或台盼蓝试验以及流式细胞术来测定该化合物对细胞增殖和细胞周期的影响。我们还使用伤口愈合迁移试验研究细胞迁移，而通过ELISA试验、蛋白质印迹法和免疫荧光显微镜来检测与细胞骨架、细胞外基质和衰老相关的蛋白质的表达。
我们发现IFC-CAF不促进增殖，但能以时间和剂量依赖的方式诱导HaCaT细胞、HDF细胞和SHDF细胞迁移；使细胞骨架蛋白（F-肌动蛋白和波形蛋白）组织更良好，并促进细胞外成分（纤连蛋白、胶原蛋白1和基质金属蛋白酶）的产生以及与细胞-底物黏着斑蛋白的黏附。IFC-CAF还能预防皮肤衰老。该处理降低了SHDF细胞中与衰老相关的标志物β-半乳糖苷酶、p53和p16INK4的表达，并提高了HaCaT细胞在UVB照射后的细胞存活率和核修复能力。
IFC-CAF具有再生特性并能抵御衰老因素，因此是一种治疗或预防皮肤衰老的潜在治疗剂。
