黑枸杞花青素、EGCG对中波紫外线诱导HaCaT细胞氧化损伤的协同保护作用

Synergistic Protective Effects of the Anthocyanins from Black Wolfberry and EGCG against the UVB-induced Oxidative Damage in HaCaT Cells

该研究探讨黑枸杞花青素(BWA)、表没食子儿茶素没食子酸酯(EGCG)的体外抗氧化能力并通过构建中波紫外线(UVB)诱导永生化人角质(HaCaT)细胞氧化损伤模型考察两者复配物对细胞氧化损伤的协同保护作用。体外抗氧化实验表明BWA、EGCG对DPPH自由基、羟自由基均具较强的清除能力和铁离子还原能力。通过Isobologram分析得出在药物安全剂量范围内,不同比例组合物对受损细胞的保护作用均优于单一物质(P<0.05),其中质量比6:4(BWA:EGCG)复配物具最强协同保护作用,协同率达17.08%,受损细胞存活率达89.71%。相较于模型组,6:4复配物使细胞内过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPX)活性分别增强2.66、1.91、2.33倍,丙二醛(MDA)含量水平降低40%(P<0.05)。结果表明BWA、EGCG均具强体外抗氧化能力,两者复配物对细胞氧化损伤具协同保护作用,其机制可能通过增强细胞中抗氧化酶活性从而减轻UVB对HaCaT细胞的氧化损伤实现,可为黑枸杞花青素与EGCG组合物的开发和应用提供理论参考及实验支撑。

In this study,the in vitro antioxidant capacities of the anthocyanin from black wolfberry(BWA)and epigallocatechin gallate(EGCG)were investigated,and the synergistically protective effect of the combination of the two compounds against the oxidative damage of immortalized human keratinocytes(HaCaT)was investigated through constructing a model of oxidative damage induced by UVB.In vitro antioxidant experiments showed that BWA and EGCG had strong DPPH radical scavenging ability, hydroxyl radical scavenging ability and iron ion reducing ability. Isobologram analysis revealed that within the safe dose range, the combinations with different proportions of BWA and EGCG exhibited greater protective effects on the damaged cells compared with BWA or EGCG alone (P<0.05), and the combination with the mass ratio of 6:4 (BWA:EGCG) exerted the strongest synergistic protection, with the synergistic rate being 17.08% and the survival rate of the damaged cells being 89.71%. Compared with the model group, the 6:4 combination increased the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) by 2.66, 1.91 and 2.33 times, respectively, while decreasing the content of malondialdehyde (MDA) by 40% (P<0.05). The results showed that both BWA and EGCG had in vitro strong antioxidant capacities, and their combination exhibited synergistically protective effect against cell oxidative damage. The mechanism might be realized by enhancing the activities of antioxidant enzymes in cells, thereby reducing UVB-induced oxidative damage to HaCaT cells, which may provide a theoretical reference and experimental support for the development and application of the combination of the anthocyanin from black wolfberry and EGCG.