探討沒食子酸還原金奈米粒子對高糖誘導人類真皮細胞中基質金屬蛋白酶表現之影響

Effect of gallic acid-reduced gold nanoparticles on high-glucose-induced the expression of matrix metalloproteinases in human dermal fibroblasts

吳昀蓁(Yun-Zhen Wu)

指導教授 ： 陳瑩容

高雄醫學大學/藥學院/香粧品學系碩士班/碩士(2019年)

摘要

高糖導致皮膚老化係透過細胞內活性氧物種 (ROS)誘導的基質金屬蛋白酶 (MMPs)表現上調，進而降解細胞外基質中之膠原蛋白。過去研究證實，金奈米粒子可抑制基質金屬蛋白酶表現，然而常規的金奈米粒子合成方法中使用之還原劑會影響環境永續性，近年來環保意識抬頭，綠色化學合成法已逐漸受到重視。本研究目的為天然抗氧化劑沒食子酸 (GA)作為還原劑與保護劑來製備金奈米粒子 (AuNPs)，並評估金奈米粒子對高糖誘導人類真皮纖維母細胞中基質金屬蛋白酶之抑制作用。在高糖誘導老化之實驗中，我們發現高糖會誘導ROS表現並激活MAPKs，透過MAPKs活化轉錄因子AP-1以及NFB來促進MMP-1的表現。我們成功利用天然抗氧化劑沒食子酸合成金奈米粒子，且金奈米粒子具有低細胞毒性並抑制高糖誘導MMP-1表現。值得注意的是，金奈米粒子能夠降低高糖所誘導產生之ROS，同時抑制ROS下游訊息ASK1表現，使ASK1下游路徑p38 MAPK與ERK皆受到抑制。金奈米粒子透過抑制MAPK減少AP-1轉錄活性，同時也降低磷酸化NFB表現，進而減少MMP-1 mRNA表現。高糖誘導MMP-1表現使第一型膠原蛋白降解，金奈米粒子能夠抑制高糖對膠原蛋白的降解作用。本研究結果提供高糖對人類真皮纖維母細胞中MMP-1調控路徑的了解，合成的金奈米粒子有效抑制MMP-1表現並降低第一型膠原蛋白降解，提供新的策略用以改善皮膚老化。

關鍵字

沒食子酸 ； 金奈米粒子 ； 抗老化 ； 綠色合成 ； 高糖

並列摘要

The mechanisms underlying high glucose-induced skin aging include the up-regulation of metalloproteinases (MMPs) activity, leading to the degradation of collagen. Some studies showed that gold nanoparticles (AuNPs) could inhibit the expression of MMPs. However, conventional methods typically involve the use of toxic compounds that are not appropriate for long-term environmental sustainability. Recently, the so-call green synthesis of AuNPs (involving biological compounds as reducing agents) has been rapidly replacing chemical methods using toxic chemicals. Gallic acid (GA) is a phenolic compound and distributed widely in nature. Thus, the effect of gallic acid-reduced gold nanoparticles (AuNPs) on MMP-1 expression in high glucose-treated human dermal fibroblasts was analyzed in the present study. We found that high glucose-induced MMP-1 expression was characteristic by increasing in ROS generation, activation of p38 MAPK/ERK and, the transcriptional activity of AP-1 and NFB. The gallic acid-reduced AuNPs had low cytotoxicity and inhibited high glucose-induced MMP-1 expression through suppression of MAPKs/AP-1 and NFB activity. Moreover, AuNPs could reduce high glucose-induced ROS generation, which suppressed ASK1 phosphorylation, and ASK1-regulated p38 MAPK and ERK activation. High glucose-induced MMP-1 expression caused degradation of type I collagen. Gallic acid-reduced AuNPs inhibited collagen degradation in high glucose-treated cells. Collectively, our data indicate that AuNPs abolishes high glucose-evoked collagen degradation through its suppressive effect on ASK1/MAPKs/AP-1-and NFB-modulated MMP-1 transcription, and suggest that gallic acid-reduced gold nanoparticles can be used as an alternative agent on reducing skin’s collagen degradation.

並列關鍵字

Gallic acid ； Gold nanoparticles ； Anti-aging ； Green synthesis ； High glucose.

參考文獻

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