Department of Orthopaedics, The Affiliated Southeast Hospital of Xiamen University, Orthopaedic Center of People's Liberation Army, Zhangzhou, 363000, China.
Department of Cardiac Surgery, The First Affiliated Hospital of Xiamen University, Xiamen, 361003, Fujian, China.
Neurochem Res. 2017 Dec;42(12):3548-3558. doi: 10.1007/s11064-017-2402-x. Epub 2017 Sep 20.
Necrostatin-1 (Nec-1) has been shown to inhibit necroptosis and convey a significant protective effect after spinal cord injury (SCI). This small molecule inhibitor may reduce tissue damage and restore neurological function by lessening mitochondrial injury after SCI and preserving energy homeostasis. However, the effects of Nec-1 on endoplasmic reticulum stress (ERS)-an important pathological consequence of SCI-are still not clear. The present study investigates the relationship between necroptosis and ERS in a rat model of SCI. Electron microscopy was employed to observe ultra-structural changes in the endoplasmic reticulum and mitochondria after lesioning. Real-time quantitative PCR was used to measure the mRNA levels of ERS-related pro-apoptotic molecules such as C/EBP homologous protein (CHOP), immunoglobulin-binding protein (BiP/GRP78) and X box-binding protein-1 (XBP-1). Western blot and immunofluorescence were conducted to analyze CHOP, GRP78 and XBP-1 protein expression after lesioning. Results demonstrated that applying Nec-1 in SCI reduces ultra-structural damage to the endoplasmic reticulum and mitochondria and inhibits expression of ERS-related genes and proteins after lesioning. Immunofluorescence also shows ERS-related proteins mainly expressed in the cytoplasm of nerve cells. Taken together, these results demonstrate that Nec-1 has protective effect on the endoplasmic reticulum and mitochondria and alleviates ERS after SCI.
Necrostatin-1（Nec-1）已被证明可抑制细胞坏死，并在脊髓损伤（SCI）后产生显著的保护作用。这种小分子抑制剂可以通过减轻 SCI 后线粒体损伤和维持能量稳态来减少组织损伤并恢复神经功能。然而，Nec-1 对内质网应激（ERS）的影响——SCI 的一个重要病理后果——尚不清楚。本研究探讨了 SCI 大鼠模型中细胞坏死与 ERS 之间的关系。电子显微镜用于观察内质网和线粒体在损伤后的超微结构变化。实时定量 PCR 用于测量 ERS 相关促凋亡分子的 mRNA 水平，如 C/EBP 同源蛋白（CHOP）、免疫球蛋白结合蛋白（BiP/GRP78）和 X 盒结合蛋白-1（XBP-1）。Western blot 和免疫荧光用于分析损伤后 CHOP、GRP78 和 XBP-1 蛋白的表达。结果表明，在 SCI 中应用 Nec-1 可减轻内质网和线粒体的超微结构损伤，并抑制损伤后 ERS 相关基因和蛋白的表达。免疫荧光也显示 ERS 相关蛋白主要表达在神经细胞的细胞质中。总之，这些结果表明 Nec-1 对内质网和线粒体具有保护作用，并减轻了 SCI 后的 ERS。
