Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, China 710032.
Free Radic Biol Med. 2012 Jul 15;53(2):183-93. doi: 10.1016/j.freeradbiomed.2012.04.015. Epub 2012 May 6.
The removal of H(2)O(2) by antioxidants has been proven to be beneficial to patients with vitiligo. Baicalein (5,6,7-trihydroxyflavone; BE) has antioxidant activity and has been used in vitiligo therapy in Chinese traditional medicine. In this study, we investigated the potential protective effect and mechanisms of BE against H(2)O(2)-induced apoptosis in human melanocytes. Melanocytes from the PIG1 cell line were pretreated with different concentrations of BE for 1 h, followed by exposure to 1.0 mM H(2)O(2) for 24 h. Cell apoptosis, reactive oxygen species levels, and mitochondrial membrane potentials were evaluated by flow cytometry, and cell viability was determined by an MTT assay. The expressions of Bax, Bcl-2, caspase-3, total and phosphorylated ERKs, and p38 MAPK were assayed by Western blot to investigate the possible molecular mechanisms. Our results showed that BE significantly inhibited H(2)O(2)-induced apoptosis, intracellular reactive oxygen species generation, and changes in the mitochondrial membrane potential. It also reduced the Bax/Bcl-2 ratio, the release of cytochrome c, the activation of caspase-3, and the phosphorylation of p38 MAPK in a concentration-dependent manner. The results demonstrate for the first time that BE exerts a cytoprotective role in H(2)O(2)-induced apoptosis by inhibiting the mitochondria-dependent caspase activation and p38 MAPK pathway.
抗氧化剂清除 H(2)O(2)已被证明对白癜风患者有益。黄芩素(5,6,7-三羟基黄酮;BE)具有抗氧化活性,并已在中药治疗白癜风中得到应用。在这项研究中,我们研究了 BE 对 H(2)O(2)诱导的人黑素细胞凋亡的潜在保护作用和机制。用不同浓度的 BE 预处理 PIG1 细胞系的黑素细胞 1 h,然后用 1.0 mM H(2)O(2)处理 24 h。通过流式细胞术评估细胞凋亡、活性氧水平和线粒体膜电位,通过 MTT 测定法确定细胞活力。通过 Western blot 测定 Bax、Bcl-2、caspase-3、总和磷酸化 ERK 和 p38 MAPK 的表达,以研究可能的分子机制。我们的结果表明,BE 显著抑制 H(2)O(2)诱导的凋亡、细胞内活性氧生成和线粒体膜电位变化。它还以浓度依赖的方式降低 Bax/Bcl-2 比值、细胞色素 c 的释放、caspase-3 的激活和 p38 MAPK 的磷酸化。结果首次表明,BE 通过抑制线粒体依赖性 caspase 激活和 p38 MAPK 途径发挥对 H(2)O(2)诱导的凋亡的细胞保护作用。