Brenneisen P, Briviba K, Wlaschek M, Wenk J, Scharffetter-Kochanek K
Dermatologische Klinik, Universität zu Köln, Germany.
Free Radic Biol Med. 1997;22(3):515-24. doi: 10.1016/s0891-5849(96)00404-2.
Reactive oxygen species (ROS) have been shown to be important messenger molecules in the induction of several genes. In human dermal fibroblasts the herbicide paraquat (PQ2+) was used to induce intracellular oxidative stress that was modulated by the inhibition of copper, zinc superoxide dismutase (Cu,ZnSOD), glutathione peroxidase (GSHPx), catalase, and blocking of the Fenton reaction. Interstitial collagenase (MMP-1) mRNA increased time dependently for up to 72 h following paraquat treatment. A correlation with the translation of MMP-1 could, however, only be detected up to 24 h, indicating an uncoupling of transcription and translation. Interleukin-1 alpha and beta mRNA showed two peaks at 6 h and 72 h. The inhibition of catalase by aminotriazol (ATZ), inhibition of GSHPx by buthionine sulfoximine (BSO), and blocking the Fenton reaction by the iron chelator desferrioxamine (DFO) in concert led to an increase in steady-state MMP-1 mRNA levels, possibly dependent on intracellular H2O2 increase. This combined treatment potentiated MMP-1 mRNA induction up to 6.5-fold compared to paraquat treated controls. Furthermore, exogenously added H2O2 caused an increase in MMP-1 mRNA levels. In contrast, inhibition of Cu,ZnSOD by diethyldithiocarbamate (DDC), leading to diminished H2O2 production from O2.-, decreased MMP-1 mRNA induction. Collectively, our data provide evidence that H2O2 is an important intermediate in the downstream signalling pathway finally leading to the induction of increased steady state MMP-1 mRNA levels. The synthesis of MMPs may contribute to connective tissue damage in vivo related to photoaging, inflammatory diseases, and tumor invasion.
活性氧（ROS）已被证明是诱导多种基因表达的重要信使分子。在人皮肤成纤维细胞中，除草剂百草枯（PQ2+）被用于诱导细胞内氧化应激，这种应激可通过抑制铜锌超氧化物歧化酶（Cu,ZnSOD）、谷胱甘肽过氧化物酶（GSHPx）、过氧化氢酶以及阻断芬顿反应来调节。百草枯处理后，间质胶原酶（MMP-1）mRNA在长达72小时内呈时间依赖性增加。然而，仅在24小时内检测到与MMP-1翻译的相关性，这表明转录和翻译发生了解偶联。白细胞介素-1α和β mRNA在6小时和72小时出现两个峰值。氨基三唑（ATZ）抑制过氧化氢酶、丁硫氨酸亚砜胺（BSO）抑制GSHPx以及铁螯合剂去铁胺（DFO）阻断芬顿反应共同导致稳态MMP-1 mRNA水平升高，这可能依赖于细胞内H2O2的增加。与百草枯处理的对照组相比，这种联合处理使MMP-1 mRNA诱导增强了6.5倍。此外，外源添加H2O2导致MMP-1 mRNA水平升高。相反，二乙基二硫代氨基甲酸盐（DDC）抑制Cu,ZnSOD导致O2.-产生的H2O2减少，从而降低了MMP-1 mRNA的诱导。总体而言，我们的数据表明H2O2是下游信号通路中的重要中间体，最终导致稳态MMP-1 mRNA水平升高。MMPs的合成可能在体内导致与光老化、炎症性疾病和肿瘤侵袭相关的结缔组织损伤。
