Sohel Md Mahmodul Hasan, Konca Yusuf, Akyuz Bilal, Arslan Korhan, Sariozkan Serpil, Cinar Mehmet Ulas
Genome and Stem Cell Centre, Erciyes University, Kayseri 38039, Turkey; Department of Animal Science, Faculty of Agriculture, Erciyes University, Kayseri 38039, Turkey.
Department of Animal Science, Faculty of Agriculture, Erciyes University, Kayseri 38039, Turkey.
Theriogenology. 2017 Jul 15;97:17-26. doi: 10.1016/j.theriogenology.2017.04.015. Epub 2017 Apr 12.
Sulforaphane (SFN) has received a great deal of research attention because of its ability to induce the production of a battery of antioxidant enzymes in certain concentrations through the activation of the Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) signaling pathway, which may effectively neutralize reactive oxygen species (ROS) induced oxidative stress. This study was conducted to investigate the potential of different concentrations of SFN in inducing antioxidative and apoptotic effects in granulosa cells (GCs). For this purpose, bovine GCs were collected from preovulatory antral follicles and cultured with different concentrations of SFN (0-80 μM) and based on phenotypic evaluation three concentrations were selected: 2 μM (low), 10 μM (medium), and 20 μM (high) for further investigations. The results showed that there was a dramatic loss of cell viability and higher cytotoxic effects of SFN on GCs at higher concentrations (>15 μM). The expression of NRF2 increased significantly (p < 0.05) with fold change ranged 3-8 in SFN treated GCs, whereas Kelch Like ECH Associated Protein 1 (KEAP1) expression was either downregulated or similar as control group under the same conditions. Moreover, the relative expression of the genes (PRDX1, CAT, TXN1and SOD1) downstream to NRF2 activation was found to be highly expressed (fold change ranged from 2 to 5, p < 0.05) in SFN treated GCs compared to the untreated control. In addition, ROS accumulation was higher in GCs treated with 20 μM SFN which in turn results in a higher accumulation of lipid droplets. Compared to control, no changes in the mitochondrial activity was observed at 2 and 10 μM SFN concentrations; however, significantly lower mitochondrial activity was found at high concentration (20 μM). The results of this study clearly showed that 10 μM SFN concentration played a crucial role in activating Nrf2 pathway without inducing apoptotic characteristics and this concentration may have beneficial effects in boosting the production of phase II antioxidant enzymes in GCs. However, at high concentration (20 μM), SFN may generate excessive ROS that causes mitochondrial dysfunction and induces cellular stress and eventually leads to apoptosis. These data strongly suggest a concentration dependent antioxidative and apoptotic effects of SFN on GCs.
萝卜硫素（SFN）因其能够通过激活核因子（红系衍生2）样2（Nrf2）信号通路，在特定浓度下诱导一系列抗氧化酶的产生而受到了大量的研究关注，这可能有效地中和活性氧（ROS）诱导的氧化应激。本研究旨在探讨不同浓度的SFN在诱导颗粒细胞（GCs）抗氧化和凋亡效应方面的潜力。为此，从排卵前的窦卵泡中收集牛颗粒细胞，并用不同浓度的SFN（0 - 80μM）进行培养，基于表型评估选择了三个浓度：2μM（低）、10μM（中）和20μM（高）用于进一步研究。结果表明，在较高浓度（>15μM）时，细胞活力显著丧失，且SFN对颗粒细胞具有更高的细胞毒性作用。在经SFN处理的颗粒细胞中，NRF2的表达显著增加（p < 0.05），倍数变化范围为3 - 8，而在相同条件下，Kelch样ECH相关蛋白1（KEAP1）的表达要么下调，要么与对照组相似。此外，与未处理的对照组相比，在经SFN处理的颗粒细胞中，发现NRF2激活下游的基因（PRDX1、CAT、TXN1和SOD1）的相对表达高度上调（倍数变化范围为2至5，p < 0.05）。此外，用20μM SFN处理的颗粒细胞中ROS积累更高，这进而导致脂滴积累增加。与对照组相比，在2和10μM SFN浓度下未观察到线粒体活性的变化；然而，在高浓度（20μM）时发现线粒体活性显著降低。本研究结果清楚地表明，10μM SFN浓度在激活Nrf2通路而不诱导凋亡特征方面起着关键作用，并且该浓度可能在促进颗粒细胞中II期抗氧化酶的产生方面具有有益作用。然而，在高浓度（20μM）时，SFN可能产生过量的ROS，导致线粒体功能障碍并诱导细胞应激，最终导致细胞凋亡。这些数据有力地表明了SFN对颗粒细胞具有浓度依赖性的抗氧化和凋亡作用。
