Department of Dermatology, The First Affiliated Hospital of Kunming Medical University, No. 295, Xichang Road, Kunming, 650032, Yunnan Province, People's Republic of China.
Lasers Med Sci. 2022 Feb;37(1):205-214. doi: 10.1007/s10103-020-03214-9. Epub 2021 Jan 5.
Due to the increase of the world's population aging, how to restore youthfulness to the skin has attracted much attention. It is well known that collagen synthesis and changes in skin barrier play an important role in the process of skin aging. However, whether Q-switched 1064-nm Nd:YAG laser (1064-QSNYL) determines the involvement of miRNAs in skin collagen synthesis and skin barrier changes remains to be elucidated. Upstream miRNAs of p38 molecular pathway have been predicted by bioinformatic database and the relationship between miRNAs and p38 verified by dual-luciferase reporter gene and Western blotting. RT-qPCR analysis detected the expression of miR-24-3p and mRNA for collagen and skin barrier-related molecules, such as keratin 10 (K10), filaggrin, and Aquaporin 4 (APQ4), in mice back skin and in the keratinocyte cell line HaCaT. Western blotting and immunofluorescence (IF) have been used to detect collagen expression and to localize, as well as quantify K10, filaggrin, and APQ4, respectively. In this study, we show that p38 is the main target gene of miRNA-24-3p, and laser irradiation at 1.5 J/cm inhibits miR-24-3p expression. Irradiation treatment upregulates the moisture, elasticity, hydroxyproline, and superoxide dismutase content of mice skin, as well as inhibits trans-epidermal water loss. Irradiation also increases collagen, K10, filaggrin, and APQ4 in both mice skin and HaCaT cells. Interestingly, we found that miR-24-3p overexpression inhibits the effect of irradiation on collagen synthesis and skin barrier. We show for the first time that 1064-QSNYL promotes collagen synthesis and protective effects on skin barrier by downregulating miR-24-3p.
由于世界人口老龄化的增加,如何使皮肤恢复年轻状态引起了广泛关注。众所周知,胶原蛋白的合成和皮肤屏障的变化在皮肤衰老过程中起着重要作用。然而,Q 开关 1064nmNd:YAG 激光(1064-QSNYL)是否决定了 miRNA 参与皮肤胶原蛋白合成和皮肤屏障变化仍有待阐明。通过生物信息学数据库预测 p38 分子途径的上游 miRNA,并通过双荧光素酶报告基因和 Western blot 验证 miRNA 与 p38 的关系。RT-qPCR 分析检测了小鼠背部皮肤和角质形成细胞系 HaCaT 中 miR-24-3p 的表达以及与胶原蛋白和皮肤屏障相关的分子(如角蛋白 10(K10)、丝聚合蛋白和水通道蛋白 4(APQ4)的 mRNA。Western blot 和免疫荧光(IF)分别用于检测胶原蛋白的表达以及 K10、丝聚合蛋白和 APQ4 的定位和定量。在这项研究中,我们表明 p38 是 miRNA-24-3p 的主要靶基因,1.5 J/cm2 的激光照射抑制了 miR-24-3p 的表达。照射处理可增加小鼠皮肤的水分、弹性、羟脯氨酸和超氧化物歧化酶含量,并抑制经表皮水分流失。照射还可增加小鼠皮肤和 HaCaT 细胞中的胶原蛋白、K10、丝聚合蛋白和 APQ4。有趣的是,我们发现 miR-24-3p 过表达抑制了照射对胶原蛋白合成和皮肤屏障的作用。我们首次表明,1064-QSNYL 通过下调 miR-24-3p 促进胶原蛋白合成和对皮肤屏障的保护作用。