Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, China.
Department of Microbiology, Xiangya School of Medicine, Central South University, Changsha, China.
Biomed Res Int. 2018 Sep 30;2018:2070971. doi: 10.1155/2018/2070971. eCollection 2018.
Oxidative stress induced by hypoxia/ischemia resulted in the excessive reactive oxygen species (ROS) and the relative inadequate antioxidants. As the initial barrier to environmental pollutants and allergic stimuli, airway epithelial cell is vulnerable to oxidative stress. In recent years, the antioxidant effect of hydrogen sulfide (HS) has attracted much attention. Therefore, in this study, we explored the impact of HS on CoCl-induced cell injury in 16HBE14o- cells. The effect of CoCl on the cell viability was detected by Cell Counting Kit (CCK-8) and the level of ROS in 16HBE14o- cells in response to varying doses (100-1000 mol/L) of CoCl (a common chemical mimic of hypoxia) was measured by using fluorescent probe DCFH-DA. It was shown that, in 16HBE14o- cells, CoCl acutely increased the ROS content in a dose-dependent manner, and the increased ROS was inhibited by the NaHS (as a donor of HS). Moreover, the calcium ion fluorescence probe Fura-2/AM and fluorescence dye Rh123 were used to investigate the intracellular calcium concentration ([Ca]) and mitochondria membrane potential (MMP) in 16HBE14o- cells, respectively. In addition, we examined apoptosis of 16HBE14o- cells with Hoechst 33342. The results showed that the CoCl effectively elevated the Ca influx, declined the MMP, and aggravated apoptosis, which were abrogated by NaHS. These results demonstrate that HS could attenuate CoCl-induced hypoxia injury via reducing ROS to perform an agonistic role for the Ca influx and MMP dissipation.
缺氧/缺血引起的氧化应激导致活性氧（ROS）过多和相对不足的抗氧化剂。作为环境污染物和过敏刺激物的初始屏障，气道上皮细胞容易受到氧化应激的影响。近年来，硫化氢（HS）的抗氧化作用引起了广泛关注。因此，在这项研究中，我们探讨了 HS 对 CoCl 诱导的 16HBE14o-细胞损伤的影响。通过细胞计数试剂盒（CCK-8）检测 CoCl 对细胞活力的影响，并用荧光探针 DCFH-DA 检测不同剂量（100-1000 mol/L）CoCl（缺氧的常见化学模拟物）对 16HBE14o-细胞中 ROS 水平的影响。结果表明，在 16HBE14o-细胞中，CoCl 急性地、剂量依赖性地增加了 ROS 含量，而 NaHS（HS 的供体）抑制了增加的 ROS。此外，使用钙离子荧光探针 Fura-2/AM 和荧光染料 Rh123 分别研究了 16HBE14o-细胞中的细胞内钙离子浓度 ([Ca]) 和线粒体膜电位 (MMP)。此外，我们用 Hoechst 33342 检测了 16HBE14o-细胞的凋亡。结果表明，CoCl 有效地增加了 Ca 内流，降低了 MMP，并加重了凋亡，而这些作用均被 NaHS 所消除。这些结果表明，HS 通过减少 ROS 来减轻 CoCl 诱导的缺氧损伤，从而发挥促进 Ca 内流和 MMP 耗散的作用。
