Department of Pathology, Chongqing Medical University, Chongqing, People's Republic of China.
J Neurosci Res. 2013 Jun;91(6):861-70. doi: 10.1002/jnr.23218. Epub 2013 Mar 29.
Oxidative stress results in protein oxidation and is implicated in cerebral disease, such as Parkinson's disease, Alzheimer's disease, and ischemic stroke. Sulfiredoxin-1 (Srxn1) is an endogenous antioxidant protein that has neuroprotective effects. The mechanisms of Srxn1 in oxidative stress have not been well studied, however. This study used 180 μM H2 O2 exposure for 24 hr to model oxidative stress. This experimental design allowed us to explore the protective effects and underlying mechanisms of Srxn1 in PC12 cells. To investigate Srxn1's role in oxidative stress protection, transient knockdowns of Srxn1 in PC12 cells were performed prior to treatment with 180 μM H2 O2 for 24 hr. Knockdown of Srxn1 resulted in decreased cell viability and increased cellular damage as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and lactate dehyrogenase analysis, respectively. Intracellular superoxide dismutase and glutathione are important indexes of oxidative stress; these were reduced in Srxn1 knockdown PC12. We further found that the decreased Srxn1 correlated with a reduction in 2-Cys Prdxs activity. Moreover, 2-Cys Prdxs protein levels were increased in the H2 O2 -dosed cells, as measured by RT-PCR and immunoblot analysis. These results suggested that Srxn1 can protect PC12 cells from H2 O2 -induced oxidative stress and are involve in Prdxs activity. Srxn1 play a protective role against oxidative injury and demonstrates potential as a target for neuroprotective intervention in oxidative stress.
氧化应激导致蛋白质氧化，并与脑部疾病（如帕金森病、阿尔茨海默病和缺血性中风）有关。硫氧还蛋白-1（Srxn1）是一种内源性抗氧化蛋白，具有神经保护作用。然而，Srxn1 在氧化应激中的作用机制尚未得到很好的研究。本研究使用 180μM H2O2 暴露 24 小时来模拟氧化应激。这种实验设计使我们能够探索 Srxn1 在 PC12 细胞中的保护作用及其潜在机制。为了研究 Srxn1 在氧化应激保护中的作用，在 180μM H2O2 处理 24 小时之前，对 PC12 细胞进行 Srxn1 的瞬时敲低。Srxn1 的敲低导致细胞活力降低，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐和乳酸脱氢酶分析分别确定细胞损伤增加。细胞内超氧化物歧化酶和谷胱甘肽是氧化应激的重要指标；Srxn1 敲低的 PC12 中这些指标降低。我们进一步发现，Srxn1 的减少与 2-Cys Prdxs 活性的降低有关。此外，通过 RT-PCR 和免疫印迹分析发现，H2O2 处理的细胞中 2-Cys Prdxs 蛋白水平增加。这些结果表明，Srxn1 可以保护 PC12 细胞免受 H2O2 诱导的氧化应激，并且与 Prdxs 活性有关。Srxn1 在氧化损伤中发挥保护作用，并显示出作为氧化应激神经保护干预的潜在目标。
