Background

Spinal cord ischemia/reperfusion injury is a common clinical, pathophysiological phenomenon with complex molecular mechanisms. Currently, there are no therapeutics available to alleviate the same. This study investigates the protective effects of sulfiredoxin-1 (Srxn 1) on spinal cord neurons following exposure to oxygen-glucose deprivation/reoxygenation (OGD/R) treatment.

Materials and methods

Primary spinal cord neurons were cultured, detected by anti-tubulin βⅢ, and transfected with adeno-associated virus (AAV)-Srxn 1 to overexpress Srxn 1. They were identified by their morphology and CCK-8 assay. The superoxide dismutase level was measured by superoxide dismutase assay. Malondialdehyde level was measured by malondialdehyde assay. The apoptosis ratio was calculated by Hoechst 33342 and Annexin V-PE/7-AAD staining. Mitochondrial transmembrane potential (Δψm) was detected by tetramethylrhodamine-methyl ester-perchlorate (TMRM) staining. The mRNA expression levels of Srxn 1 and caspase 3 were detected by quantitative reverse transcription-polymerase chain reaction, and the protein expression levels of Srxn 1, bax, bcl-2, cytosolic cytochrome c, and caspase 3 were detected by western blotting.

Results

AAV-Srxn 1 up-regulated mRNA and protein levels of Srxn 1 in spinal cord neurons. Following exposure to OGD/R, overexpression of Srxn 1 improved the neuronal viability, alleviated the neuron apoptosis, enhanced the mitochondrial transmembrane potential, increased the SOD level, decreased the MDA level, inhibited the expression of cytosolic cytochrome c, bax, and caspase 3, and promoted the expression of bcl-2.

Conclusion

Srxn 1 plays a significant role in anti-apoptosis of spinal cord neurons, and Srxn 1 may be a potential therapeutic target for spinal cord I/R injury.

中文翻译：

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Sulfiredoxin-1通过bax /细胞色素c / caspase 3凋亡途径在氧-葡萄糖剥夺/复氧模型中保护脊髓神经元免受氧化应激

背景

脊髓缺血/再灌注损伤是一种具有复杂分子机制的常见临床病理生理现象。当前，没有可用于缓解相同症状的疗法。这项研究调查了暴露于氧-葡萄糖剥夺/复氧（OGD / R）治疗后sulfiredoxin-1（Srxn 1）对脊髓神经元的保护作用。

材料和方法

培养原代脊髓神经元，用抗微管蛋白βⅢ检测，并用腺相关病毒（AAV）-Srxn 1转染以过量表达Srxn1。通过形态学和CCK-8分析鉴定它们。通过超氧化物歧化酶测定法测量超氧化物歧化酶水平。丙二醛水平通过丙二醛测定法测量。通过Hoechst 33342和膜联蛋白V-PE / 7-AAD染色计算细胞凋亡率。通过四甲基罗丹明-高氯酸甲酯（TMRM）染色检测线粒体跨膜电位（Δψm）。通过定量逆转录-聚合酶链反应检测Srxn 1和caspase 3的mRNA表达水平，并通过western印迹检测Srxn 1，bax，bcl-2，胞浆细胞色素c和caspase 3的蛋白表达水平。

结果

AAV-Srxn 1上调脊髓神经元中Srxn 1的mRNA和蛋白水平。暴露于OGD / R后，Srxn 1的过表达改善了神经元的生存能力，减轻了神经元的凋亡，增强了线粒体跨膜电位，增加了SOD的水平，降低了MDA的水平，抑制了胞浆细胞色素c，bax和caspase 3的表达。 ，并促进bcl-2的表达。

结论

Srxn 1在脊髓神经元的抗凋亡中起着重要作用，Srxn 1可能是脊髓I / R损伤的潜在治疗靶标。