(1.山西长治医学院 生物化学教研室, 山西 长治 046099;2.山西长治医学院 职业与环境卫生学教研室, 山西 长治 046099;3.山西长治医学院 化学教研室, 山西 长治 046099)
(1.Department of Biochemistry, Changzhi Medical College, Changzhi 046099, China;2.Department of Occupational and Environmental Health, Changzhi Medical College, Changzhi 046099,China;3.Department of Chemistry, Changzhi Medical College, Changzhi 046099, China)
Science and Technology Innovation Project of Universities in Shanxi Province (2019L0675).
为了研究葡萄籽粗多糖(crude polysaccharides from grape seeds,GSCPs)体外抗氧化作用及对秀丽隐杆线虫的体内抗氧化作用,采用水提醇沉法提取GSCPs,检测GSCPs对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基和羟自由基的清除作用及对DNA氧化损伤的抑制作用；建立RAW 264.7巨噬细胞氧化损伤模型,在细胞水平探讨GSCPs的抗氧化能力；同时利用秀丽隐杆线虫研究GSCPs的体内抗氧化功能。体外实验结果表明:GSCPs可有效清除自由基,抑制DNA的氧化损伤,质量浓度为0.4mg/mL的GSCPs对DPPH自由基的清除率达84%,对羟自由基清除率为89%；GSCPs可以下调H2O2诱导的RAW 264.7巨噬细胞活性氧(reactive oxygen species,ROS)水平,正向调节细胞内超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性,质量浓度为0.2mg/mL的GSCPs处理组可使细胞内SOD活性从H2O2处理组的81.1%升至96.3%,GSH-Px活性从H2O2处理组的92.1%升至99.6%。此外,GSCPs可延长秀丽隐杆线虫寿命,提高其对抗急性氧化应激的能力,有效清除秀丽隐杆线虫体内的ROS。质量浓度为0.8mg/mL GSCPs处理组秀丽隐杆线虫的平均寿命较对照组延长27.67%,将急性氧化应激的秀丽隐杆线虫平均存活时间延长33.58%,秀丽隐杆线虫体内ROS生成量较对照组可降低56.33%。因此,葡萄籽粗多糖在体内外均表现出良好的抗氧化性,可用于抗氧化功能产品的开发。
To study the antioxidant effect of crude polysaccharides from grape seeds(GSCPs) in vitro and its in vivo antioxidant activity on Caenorhabditis elegans(C.elegans), GSCPs were extracted and isolated by water extraction and alcohol precipitation. The scavenging effect of GSCPs on 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical and hydroxyl free radical as well as their inhibitory effect on DNA oxidative damage were detected in vitro. The antioxidant capacity of GSCPs on cellular level was investigated by H2O2-induced oxidative damage model of RAW 264.7 macrophages. In addition, the antioxidant function in vivo of GSCPs was studied by C.elegans. The results of in vitro experiment demonstrated that GSCPs could effectively scavenge free radicals and inhibit oxidative damage to DNA. The scavenging activities of 0.4mg/mL GSCPs for DPPH and hydroxyl radical were 84% and 89%, respectively. GSCPs could attenuate H2O2-induced oxidative damage in RAW 264.7 macrophages by decreasing reactive oxygen species(ROS) levels and increasing the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px). Compared with H2O2 treated group, the activity of SOD and GSH-Px increased from 81.1% to 96.3% and from 92.1% to 99.6%, respectively after treated with 0.2mg/mL GSCPs. GSCPs could prolong C.elegans lifespan, improve the ability to resist acute oxidative stress, and effectively remove ROS in C.elegans. Compared with the control group, the average lifespan of C. elegans and the average survival time under acute oxidative stress were prolonged by 27.67% and 33.58%, respectively in the 0.8mg/mL GSCPs treated group. In contrast, GSCPs could cause 56.33% reduction in ROS levels of C.elegans. Therefore, GSCPs showed good antioxidant activicy in vivo and in vitro, which could be used as potential antioxidant ingredients in functional foods.