Xiao Xiaoqing, Huang Minghuan, Fan Chunyan, Zuo Fuguo
Department of Dermatology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, P.R. China.
Exp Ther Med. 2021 Feb;21(2):157. doi: 10.3892/etm.2020.9588. Epub 2020 Dec 17.
Skin and in particular photoaging or premature aging, are caused by a variety of factors, including hormone imbalance and exposure to ultraviolet radiation. The aim of the present study was to explore the roles of Dual oxidase 2 (DUOX2) and related NF-κB signals in skin photoaging. Cell models of photoaging were constructed by irradiating human skin fibroblast lines (HSF2) with ultraviolet B (UVB) of different doses (0, 15, 30 and 60 mj/cm). The cell counting kit-8 (CCK8) was used to determine cell proliferation. Flow cytometry was used to determine the production of reactive oxygen species (ROS). A biochemical method was to determine the content of hydrogen peroxide, and the quantitative PCR (qPCR) was used to determine the expression of matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), Col-Ⅰ and α-SMA in the cells. Enzyme-linked immunosorbent assay (ELISA) was used to determine the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Western blot analysis was performed to determine the expression of DUOX2, p65 and p-p65. The results showed that,UVB irradiation dose- and time-dependently inhibited the proliferation of HSF2 cells. Cellular inflammatory response, ROS production and hydrogen peroxide increase was promoted. Col-Ⅰ and α-SMA were downregulated, MMP2 and MMP9 were upregulated, and the phosphorylation of NF-κB p65 was promoted. The above indicators were all reversed by interference with DUOX2. Overexpression of DUOX2 has an effect that is similar to UVB irradiation, but the effects can be significantly weakened by NF-κB inhibitor, NAC. Upregulation of DUOX2 expression plays a crucial role in UVB-induced aging of HSF2 cells. The specific mechanism is related to the promotion of ROS production and cellular inflammatory response and activation of NF-κB signals.
皮肤，尤其是光老化或过早衰老，是由多种因素引起的，包括激素失衡和紫外线辐射暴露。本研究的目的是探讨双氧化酶2（DUOX2）和相关核因子κB信号在皮肤光老化中的作用。通过用不同剂量（0、15、30和60 mj/cm）的紫外线B（UVB）照射人皮肤成纤维细胞系（HSF2）构建光老化细胞模型。使用细胞计数试剂盒-8（CCK8）测定细胞增殖。采用流式细胞术测定活性氧（ROS）的产生。采用生化方法测定过氧化氢含量，采用定量聚合酶链反应（qPCR）测定细胞中基质金属蛋白酶2（MMP2）、基质金属蛋白酶9（MMP9）、Ⅰ型胶原（Col-Ⅰ）和α-平滑肌肌动蛋白（α-SMA）的表达。采用酶联免疫吸附测定（ELISA）法测定肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的表达。进行蛋白质免疫印迹分析以测定DUOX2、p65和磷酸化p65的表达。结果表明，UVB照射剂量和时间依赖性地抑制HSF2细胞的增殖。促进了细胞炎症反应、ROS产生和过氧化氢增加。Col-Ⅰ和α-SMA下调，MMP2和MMP9上调，核因子κB p65的磷酸化被促进。上述指标均因干扰DUOX2而逆转。DUOX2的过表达具有与UVB照射相似的作用，但核因子κB抑制剂NAC可显著减弱其作用。DUOX2表达上调在UVB诱导的HSF2细胞衰老中起关键作用。具体机制与促进ROS产生、细胞炎症反应以及核因子κB信号激活有关。
