Huang Chieh-Chen, Wu Wen-Bin, Fang Jia-You, Chiang Han-Sun, Chen Shao-Kuan, Chen Bing-Huei, Chen Ying-Ting, Hung Chi-Feng
School of Medicine, Fu-Jen Catholic University, Taipei, Taiwan.
Molecules. 2007 Aug 14;12(8):1845-58. doi: 10.3390/12081845.
(-)-Epicatechin-3-gallate (ECG) is a polyphenolic compound similar to (-)-epigallocatechin-3-gallate (EGCG) which is abundant in green tea. Numerous workers have proposed that EGCG protects epidermal cells against UVB-induced damage. However, little has been known about whether ECG protects keratinocytes against UVB-induced damage. We decided to investigate the protective effects and underlying mechanisms of ECG on UVB-induced damage. Cell viability was determined by the MTT assay. Activation of ERK1/2, p38 and JNK was analyzed by Western blotting. Intracellular H2O2 production and DNA content was analyzed by flow cytometry. Lipid peroxidation was assayed by colorimetry. In our study, we found that ECG dose-dependently attenuated UVB-induced keratinocyte death. Moreover, ECG markedly inhibited UVB-induced cell membrane lipid peroxidation and H2O2 generation in keratinocytes, suggesting that ECG can act as a free radical scavenger when keratinocytes were photodamaged. In parallel, H2O2-induced the activation of ERK1/2, p38 and JNK in keratinocytes could be inhibited by ECG. UVB-induced pre-G1 arrest leading to apoptotic changes of keratinocytes were blocked by ECG. Taken together, we provide here evidence that ECG protects keratinocytes from UVB-induced photodamage and H2O2-induced oxidative stress, possibly through inhibition of the activation of ERK1/2, p38 and JNK and/or scavenging of free radicals.
(-)-表儿茶素-3-没食子酸酯(ECG)是一种多酚类化合物,与绿茶中大量存在的(-)-表没食子儿茶素-3-没食子酸酯(EGCG)类似。许多研究人员提出,EGCG可保护表皮细胞免受紫外线B(UVB)诱导的损伤。然而,关于ECG是否能保护角质形成细胞免受UVB诱导的损伤,人们所知甚少。我们决定研究ECG对UVB诱导损伤的保护作用及其潜在机制。通过MTT法测定细胞活力。通过蛋白质免疫印迹法分析细胞外信号调节激酶1/2(ERK1/2)、p38和c-Jun氨基末端激酶(JNK)的激活情况。通过流式细胞术分析细胞内过氧化氢(H2O2)的产生和DNA含量。通过比色法测定脂质过氧化。在我们的研究中,我们发现ECG能剂量依赖性地减轻UVB诱导的角质形成细胞死亡。此外,ECG显著抑制UVB诱导的角质形成细胞膜脂质过氧化和H2O2生成,这表明当角质形成细胞受到光损伤时,ECG可作为自由基清除剂发挥作用。同时,ECG可抑制H2O2诱导的角质形成细胞中ERK1/2、p38和JNK的激活。ECG可阻断UVB诱导的G1期前停滞,从而导致角质形成细胞的凋亡变化。综上所述,我们在此提供证据表明,ECG可能通过抑制ERK1/2、p38和JNK的激活和/或清除自由基,保护角质形成细胞免受UVB诱导的光损伤和H2O2诱导的氧化应激。