软枣猕猴桃原花青素对H2O2诱导细胞损伤的预保护作用

Preprotective Effect of Actinidia arguta Proanthocyanidins on the Cell Injuried by Drogen Peroxide

摘要:

研究软枣猕猴桃中原花青素类化合物对过氧化氢损伤人永生化表皮细胞（HaCat）的保护作用。以浏阳市大围山软枣猕猴桃为试材，以体外培养的HaCat细胞为实验对象。试验分为3组，分别为对照组、过氧化氢损伤组、处理组（软枣猕猴桃原花青素5~1000 μg/mL预处理），以细胞内细胞存活率（凋亡率）、SOD（超氧化物歧化酶）、GSH-PX（谷胱甘肽过氧化物酶）、ROS（活性氧簇）、Nrf2基因及其蛋白表达水平为综合评价指标，评价其抗氧化性强弱。实验结果表明：当加入原花青素类化合物对细胞进行预保护后，5~500 μg/L浓度的原花青素对损伤细胞的存活率为77%左右，较模型组提高了2.88倍。处理组细胞内SOD（10.46 U/mg prot）、GSH-PX水平（33.83 U/mg prot）较模型组升高了108.28%、137.48%，且存在明显差异(p<0.05)；而ROS水平显著下降，与模型组（96.21）相比细胞内平均荧光强度下降了48.60%。此时Nrf2基因及其蛋白表达量分别为1.40、35.11较模型组分别提高了63.93%、80.70%。这说明软枣猕猴桃中原花青素类化合物对H2O2造成HaCat细胞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有效抗氧化剂产品应用于市场中。

Abstract:

The protective effect of Actinidia arguta Proanthocyanidinss on the Human immortalized epidermal cells (HaCat) injuried by drogen peroxide was investigated. Actinidia arguta in Dawei Mountain of Liuyang was used as test material and HaCat cells cultured in vitro were used as the experimental objects. The experiment was divided into three groups: control group, hydrogen peroxide injury group and treatment group (pretreatment with 5~1000 μg/mL proanthocyanidins of Actinidia arguta), the cell survival rate (apoptosis rate), SOD (superoxide dismutase), GSH-PX (glutathione peroxidase), ROS (reactive oxygen species), Nrf2 gene and protein expression level were measured to evaluate their antioxidant activity. Results showed that when the proanthocyanidins were added to the cells to protect the cells, the survival rate of the injured cells was about 77% at the concentration of 5~500 μg/L proanthocyanidins, which was 2.88 times higher than that in the model group. The levels of SOD (10.46 U/mg prot) and GSH-PX (33.83 U/mg prot) in the treatment group were 108.28% and 137.48% higher than those in the model group (p<0.05), respectively. The level of ROS decreased significantly, and the average intracellular fluorescence intensity decreased by 48.60% compared with the model group (96.21). Meanwhile, the expression of Nrf2 and its protein were 1.40 and 35.11, respectively, which were 63.93% and 80.70% higher than that in the model group. These indicated that procyanidins in Actinidia chinensis have a preprotective effect on oxidative damage of HaCat cells induced by H2O2, and can be further developed into natural and effective antioxidant products to be used in the industry.