Institut für Pharmakologie und Klinische Pharmakologie, Universitätsklinikum Düsseldorf, 40225 Düsseldorf, Germany.
J Biol Chem. 2011 May 20;286(20):18268-76. doi: 10.1074/jbc.M110.201665. Epub 2011 Mar 17.
UVB irradiation causes characteristic features of skin aging including remodeling of the dermal extracellular matrix. A key feature during this process is the up-regulation of matrix metalloproteinases and cleavage of collagen. Hyaluronic acid (HA), a major component of the dermal matrix, decreases after chronic UVB exposure. However, the factors that govern the decline of HA synthesis during the course of actinic aging are largely unknown. The aim of the present study was to explore whether collagen degradation causes inhibition of HA synthesis in human skin fibroblasts. After treatment of fibroblasts with collagen fragments (CF) in vitro, resolution of the actin cytoskeleton and inhibition of HA secretion occurred because of specific down-regulation of hyaluronan synthase 2 (HAS2) expression. The α(v)β(3)-agonist, RGDS, latrunculin A, and an inhibitor of Rho-activated kinase inhibited HAS2 expression. Conversely, blocking antibodies to α(v)β(3) abolished the down-regulation of HAS2 and the cytoskeletal effects. Furthermore, inhibition of cofilin phosphorylation in response to CF was prevented by α(v)β(3)-blocking antibodies. The key role of ERK signaling was shown by reduced nuclear accumulation of phosphoERK and of ELK-1 phosphorylation in response to CF. In addition, the ERK inhibitor PD98059 reduced HAS2 expression. Also, UVB irradiation of fibroblasts caused down-regulation of HAS2, which was sensitive to matrix metalloproteinase inhibitors and to α(v)β(3)-blocking antibodies. In conclusion, these data suggest that CF activate α(v)β(3)-integrins and in turn inhibit Rho kinase (ROCK) signaling and nuclear translocation of phosphoERK, resulting in reduced HAS2 expression. Therefore, a novel mechanism is presented how proteolytic collagen cleavage may inhibit HA synthesis in dermal fibroblasts during extrinsic skin aging.
中波紫外线照射会导致皮肤老化的特征性变化,包括真皮细胞外基质的重塑。在这个过程中,一个关键特征是基质金属蛋白酶的上调和胶原蛋白的裂解。透明质酸(HA)是真皮基质的主要成分,在慢性中波紫外线照射后会减少。然而,在光老化过程中控制 HA 合成下降的因素在很大程度上尚不清楚。本研究旨在探讨胶原降解是否会导致人皮肤成纤维细胞中 HA 合成的抑制。体外用胶原片段(CF)处理成纤维细胞后,由于透明质酸合酶 2(HAS2)表达的特异性下调,肌动蛋白细胞骨架解聚和 HA 分泌受到抑制。α(v)β(3)-激动剂 RGDS、拉库氯铵 A 和 Rho 激活激酶抑制剂抑制 HAS2 表达。相反,阻断 α(v)β(3)抗体可消除 HAS2 的下调和细胞骨架的作用。此外,CF 引起的 cofilin 磷酸化抑制作用被α(v)β(3)阻断抗体所阻止。阻断抗体还减少了 ERK 信号的核积累。此外,ERK 抑制剂 PD98059 也降低了 HAS2 的表达。此外,中波紫外线照射还会导致 HAS2 的下调,这种下调对基质金属蛋白酶抑制剂和α(v)β(3)阻断抗体敏感。总之,这些数据表明 CF 激活了 α(v)β(3)-整联蛋白,进而抑制了 Rho 激酶(ROCK)信号和磷酸化 ERK 的核易位,导致 HAS2 表达减少。因此,提出了一种新的机制,即蛋白水解胶原裂解如何在真皮成纤维细胞的外在皮肤老化过程中抑制 HA 的合成。