Traumatic brain injury (TBI) is the primary cause of death and disability affecting over 10 million people in the industrialized world. TBI causes a wide spectrum of secondary molecular and cellular complications in the brain. However, the pathological events are still not yet fully understood. Previously, we have shown that the glia maturation factor (GMF) is a mediator of neuroinflammation in neurodegenerative diseases. To identify the potential molecular pathways accompanying TBI, we used an in vitro cell culture model of TBI. A standardized injury was induced by scalpel cut through a mixed primary cell culture of astrocytes, microglia and neurons obtained from both wild type (WT) and GMF-deficient (GMF-KO) mice. Cell culture medium and whole–cell lysates were collected at 24, 48, and 72 h after the scalpel cuts injury and probed for oxidative stress using immunofluorescence analysis. Results showed that oxidative stress markers such as glutathione and glutathione peroxidase were significantly reduced, while release of cytosolic enzyme lactate dehydrogenase along with nitric oxide and prostaglandin E2 were significantly increased in injured WT cells compared with injured GMF-KO cells. In addition, injured WT cells showed increased levels of oxidation product 4-hydroxynonenal and 8-oxo-2′-deoxyguanosine compared with injured GMF-KO cells. Further, we found that injured WT cells showed a significantly increased expression of glial fibrillary acidic protein, ionized calcium binding adaptor molecule 1, and phosphorylated ezrin/radixin/moesin proteins, and reduced microtubule associated protein expression compared with injured GMF-KO cells after injury. Collectively, our results demonstrate that GMF exacerbates the oxidative stress–mediated neuroinflammation that could be brought about by TBI-induced astroglial activation.

中文翻译：

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神经胶质成熟因子介导的神经炎症在创伤性脑损伤的体外模型中加剧了神经元损伤。

创伤性脑损伤 (TBI) 是影响工业化世界超过 1000 万人的死亡和残疾的主要原因。TBI 在大脑中引起广泛的继发性分子和细胞并发症。然而，病理事件仍未完全了解。以前，我们已经表明神经胶质成熟因子 (GMF) 是神经退行性疾病中神经炎症的介质。为了确定伴随 TBI 的潜在分子途径，我们使用了体外TBI的细胞培养模型。标准化损伤是通过手术刀切开星形胶质细胞、小胶质细胞和从野生型 (WT) 和 GMF 缺陷 (GMF-KO) 小鼠获得的神经元的混合原代细胞培养物引起的。在手术刀切割损伤后 24、48 和 72 小时收集细胞培养基和全细胞裂解物，并使用免疫荧光分析探测氧化应激。结果表明，与受损的 GMF-KO 细胞相比，受损 WT 细胞中谷胱甘肽和谷胱甘肽过氧化物酶等氧化应激标志物显着减少，而胞质酶乳酸脱氢酶以及一氧化氮和前列腺素 E2 的释放显着增加。此外，与受损的 GMF-KO 细胞相比，受损的 WT 细胞显示出更高水平的氧化产物 4-羟基壬烯醛和 8-oxo-2'-脱氧鸟苷。此外，我们发现与损伤后受损的 GMF-KO 细胞相比，受损的 WT 细胞的胶质纤维酸性蛋白、离子钙结合接头分子 1 和磷酸化的 ezrin/radixin/moesin 蛋白的表达显着增加，并降低了微管相关蛋白的表达. 总的来说，我们的结果表明 GMF 加剧了氧化应激介导的神经炎症，这可能由 TBI 诱导的星形胶质细胞激活引起。