目的:内在皮肤老化是一个不可避免的过程,在真皮-表皮连接(DEJ)中,细胞外基质沉积减少,机械完整性受损。透明质酸是最有前途的天然成分之一。在这项研究中,揭示了新型透明质酸复合物对抗内在皮肤老化的多种机制。
方法:采用免疫组织化学分析和酶联免疫吸附试验评价低分子量透明质酸钠的效果。其乙酰化衍生物和HA复合物对正常人成纤维细胞中基质金属蛋白酶-1(MMP-1)和I型胶原的表达。然后,进行了免疫组织化学分析和苏木精和伊红染色,以评估HA复合物在重建皮肤等效物中的相同效果,以及它对组织学结构和DEJ的好处。
结果:在正常人真皮成纤维细胞中,透明质酸复合物,其中含有低分子量透明质酸钠及其乙酰化衍生物,通过增加I型胶原表达具有协同作用。同时,MMP-1的产生受到抑制。这在随后的皮肤等效实验中得到了证实,有趣的是,还发现透明质酸复合物可增加两种DEJ蛋白的表达。
结论:本概念验证研究中的多机制透明质酸复合物通过抑制MMP-1的表达,增强I型胶原的积累和DEJ蛋白的表达,在体外表现出皮肤抗衰老作用。这揭示了研究各种类型透明质酸更多生物活性的新途径。
OBJECTIVE: Intrinsic skin aging is an inevitable process with reduced extracellular matrix deposition and impaired mechanical integrity in the dermal-epidermal junction (DEJ). Hyaluronan is one of the most promising natural ingredients. In this research, multiple mechanisms of a novel hyaluronan complex against intrinsic skin aging were revealed.
METHODS: Immunohistochemical analysis and enzyme-linked immunosorbent assay were employed to evaluate the effect of low-molecular weight sodium hyaluronan, its acetylated derivative and HA complex on expression of matrix metalloproteinase-1 (MMP-1) and type I collagen in normal human fibroblasts. Then, immunohistochemical analysis and hematoxylin and eosin staining was carried out to evaluate identical effects of HA complex in reconstructed skin equivalents, as well as its benefits on histological structure and DEJ.
RESULTS: In normal human dermal fibroblasts, the hyaluronan complex, which contains low-molecular weight sodium hyaluronate and its acetylated derivative, has synergistic effects by increasing type I collagen expression. At the same time, MMP-1 production was inhibited. This was confirmed in subsequent experiments with skin equivalent, and intriguingly, the hyaluronan complex was also found to increase the expression of two DEJ proteins.
CONCLUSIONS: The multimechanism hyaluronan complex in this proof-of-concept study exhibited skin antiaging effects in vitro through inhibiting the expression of MMP-1 and enhancing type I collagen accumulation and the expression of DEJ proteins, which reveals new avenues for investigating more biological activities of various types of hyaluronan.