摘要
研究软枣猕猴桃中黄酮类化合物对过氧化氢(H_2O_2)损伤人永生化表皮细胞(HaCaT细胞)的保护作用。以湖南省浏阳市大围山软枣猕猴桃为实验材料,以体外培养的HaCaT细胞为实验对象。实验分为空白对照组、H_2O_2模型组、阳性对照组(VC组)和黄酮处理组(软枣猕猴桃黄酮0.1~1.0 mg/mL预处理),以细胞内超氧化物歧化酶(superoxide dismutase,SOD)活力、活性氧簇(reactive oxygen species,ROS)水平为综合评价指标,并以VC作为阳性对照评价其抗氧化活性。实验结果表明:当H_2O_2浓度为30μmol/L时,细胞相对存活率为40%左右,较好地诱导了HaCaT细胞氧化损伤模型。当加入黄酮类化合物对细胞进行预保护后,质量浓度0.3、0.6 mg/mL的黄酮处理组细胞相对存活率为60%左右,较模型组均提高了20%左右。0.3 mg/mL黄酮处理组细胞内SOD活力(6.33 U/mg)较模型组升高了约1倍,且存在明显差异(P<0.05);ROS水平显著下降,与模型组(677.80)相比下降了13%左右。说明软枣猕猴桃中黄酮类化合物对H_2O_2造成的氧化损伤具有一定的预保护作用,可进一步将其开发成相应的天然抗氧化剂产品投入市场。
The protective effect of Actinidia arguta flavonoids(AAF) on H_2O_2-induced oxidative injury in human immortalized epidermal cells(HaCaT) was investigated in this study. The flavonoids were extracted from A. arguta grown in Da Wei Mountain, Liuyang, Hunan provice. Ha Ca T cells were cultured in vitro. The experiment was divided into control, hydrogen peroxide damage, positive control(VC), and AAF treatment groups(at concentrations of 0.1–1.0 mg/m L). Superoxide dismutase(SOD) activity and reactive oxygen species(ROS) levels in cells were measured to evaluate the cytoprotective effect of AAF. VC was used as a positive control to evaluate its antioxidant activity. The results showed that at a H_2O_2 concentration of 30 μmol/L, the survival rate of the cells was approximately 40% and the oxidative damage model was established. When AAF at concentrations of 0.3 and 0.6 mg/mL was used to protect the cells, the survival rate was about 60%, which increased by about 20% compared with the model group. SOD activity(6.33 U/mg) in the treatment group(0.3 mg/m L) was about 2 times higher than that in the model group, with a significant difference(P < 0.05). ROS level decreased significantly by 13% when compared with the model group(677.80). These data suggested a protective effect of AAF on H_2O_2-indued oxidative damage. Therefore, it can be further developed into a potent natural antioxidant product.
引文
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