为探究雨生红球藻源虾青素(Haematococcus pluvialis ester astaxanthin, E-AST)和人工合成虾青素(synthetic astaxanthin, S-AST)抗氧化能力的差异,该研究分析了不同浓度(0、2.5、5、10 μmol/L)2种来源虾青素处理24 h后,对HepG2细胞存活率、细胞内不同活性氧分子含量和细胞内抗氧化活性(cellular antioxidant activity, CAA)的影响。结果显示,与对照组相比,E-AST和S-AST对HepG2细胞存活率无明显影响,仅10 μmol/L E-AST能够显著提高细胞存活率。荧光探针DCFH-DA检测显示,与对照组比较,除5 μmol/L E-AST和2.5 μmol/L S-AST外,添加其他浓度的2种虾青素均能够显著降低细胞内活性氧(reactive oxygen species, ROS)含量(P<0.05),但是同一浓度2种虾青素处理组之间不存在显著性差异(P>0.05)。DHR123荧光探针的检测结果则显示,在虾青素处理浓度为5 μmol/L时,E-AST对H2O2的清除能力显著高于S-AST(P<0.05)。而使用DHE探针检测发现,仅2.5 μmol/L S-AST处理后细胞内·O2-的含量显著降低(P<0.05),其他处理组与对照组比略有减少但不存在显著性差异(P>0.05)。2种虾青素的CAA值随虾青素处理浓度的升高而升高,但同一浓度不同来源虾青素之间CAA值无显著性差异(P>0.05)。综上所述,2种不同来源虾青素抗氧化能力存在差异,并可能与2种虾青素对HepG2细胞内活性氧分子的清除能力不同有关。
To explore the difference in antioxidant capacity between astaxanthin from Haematococcus pluvialis (E-AST) and synthetic astaxanthin (S-AST), the effects of astaxanthin on cell viability, different active oxygen molecules, and cellular antioxidant activity (CAA) in HepG2 cells were analyzed after 24 h treatment. Results showed that E-AST and S-AST had no obvious toxic effects on HepG2 cells, and 10 μmol/L E-AST could significantly improve the viability of HepG2 cells. Fluorescence probe DCFH-DA identified that both of E-AST and S-AST could significantly decrease ROS content in HepG2 cells compared with the control group (P<0.05), except 5 μmol/L E-AST and 2.5 μmol/L S-AST. However, there was no significant difference between the two astaxanthin treatment groups with the same concentration (P>0.05). The detection results with DHR123 showed that the scavenging capacity of 5 μmol/L E-AST on hydrogen peroxide (H2O2) was significantly higher than that of S-AST (P<0.05). Furthermore, superoxide anion free radical (·O2-) specific probes DHE demonstrated that the ·O2- content decreased significantly exposed to 2.5 μmol/L S-AST (P<0.05), while other treatment groups had no significant change compared with the control group (P>0.05). The CAA of the two astaxanthins was increased does-dependent, but there was no significant difference between E-AST and S-AST at the same concentration (P>0.05). These results suggest that the antioxidant capacity of astaxanthin from two different sources is different in HepG2 cells by scavenging reactive oxygen species.
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