Xiang Ya, Lai Fangnong, He Guifang, Li Yapeng, Yang Leilei, Shen Wei, Huo Heqiang, Zhu Jun, Dai Hongyi, Zhang Yugang
College of Horticulture, Qingdao Agricultural University, Qingdao, China.
Qingdao Key Laboratory of Genetic Development and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, China.
PLoS One. 2017 Aug 29;12(8):e0184033. doi: 10.1371/journal.pone.0184033. eCollection 2017.
Anthocyanins are the polyphenolic phytochemicals which have been shown to scavenge free radicals. In this study, we investigated the effects of anthocyanins extracted from red-fleshed apples (Malus sieversii) on reducing oxidative damage by Rosup in porcine granulosa cells (GCs) by measuring intracellular reactive oxygen species (ROS), content of glutathione (GSH), activities of superoxide dismutase (SOD1), catalase (CAT) and glutathione peroxidase (GPX1) and the gene expression of SOD1, CAT, GPX1. Apoptosis was determined with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and apoptosis-related proteins were quantified with Western blotting. The results indicate that Rosup increases oxidative stress by inducing reactive oxygen species production in porcine GCs and the oxidative stress could be reduced by anthocyanins. The gene expression of SOD1, CAT, GPX1 and the activities of these enzymes were increased when GCs were treated with anthocyanins and Rosup for 6 hours. Anthocyanins inhibit Rosup-induced apoptosis by increasing expression of antiapoptotic protein Bcl-2 and suppressing the expression of pro-apoptotic protein Bax. Collectively, anthocyanins from red-fleshed apples reduce oxidative stress and inhibit apoptosis in porcine GCs in vitro. This approach indicates that antioxidants might be developed from red-fleshed apples.
花青素是一类已被证明能清除自由基的多酚类植物化学物质。在本研究中，我们通过测量细胞内活性氧（ROS）、谷胱甘肽（GSH）含量、超氧化物歧化酶（SOD1）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GPX1）的活性以及SOD1、CAT、GPX1的基因表达，研究了从红肉苹果（新疆野苹果）中提取的花青素对降低猪颗粒细胞（GCs）中由Rosup诱导的氧化损伤的影响。采用TdT介导的dUTP生物素缺口末端标记法（TUNEL）测定细胞凋亡，并通过蛋白质免疫印迹法对凋亡相关蛋白进行定量分析。结果表明，Rosup通过诱导猪颗粒细胞产生活性氧来增加氧化应激，而花青素可以减轻这种氧化应激。当颗粒细胞用花青素和Rosup处理6小时后，SOD1、CAT、GPX1的基因表达以及这些酶的活性均增加。花青素通过增加抗凋亡蛋白Bcl-2的表达和抑制促凋亡蛋白Bax的表达来抑制Rosup诱导的细胞凋亡。总体而言，红肉苹果中的花青素可减轻体外培养的猪颗粒细胞的氧化应激并抑制细胞凋亡。该研究表明红肉苹果有望开发成为抗氧化剂。
