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Neurotoxic role of interleukin-17 in neural stem cell differentiation after intracerebral hemorrhage

机译：白细胞介素-17在脑出血后神经介素-17中的神经毒素作用

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Interleukin 17 (IL-17) and its main producer, T cell receptor γδ cells, have neurotoxic effects in the pathogenesis of intracerebral hemorrhage (ICH), aggravating brain injuries. To investigate the correlation between IL-17 and ICH, we dynamically screened serum IL-17 concentrations using enzyme-linked immunosorbent assay and explored the clinical values of IL-17 in ICH patients. There was a significant negative correlation between serum IL-17 level and neurological recovery status in ICH patients (r = –0.498, P 0.01). To study the neurotoxic role of IL-17, C57BL/6 mice were used to establish an ICH model by injecting autologous blood into the caudate nucleus. Subsequently, the mice were treated with mouse neural stem cells (NSCs) and/or IL-17 neutralizing antibody for 72 hours. Flow cytometry, brain water content detection, Nissl staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling results indicated that NSC transplantation significantly reduced IL-17 expression in peri-hematoma tissue, but there was no difference in T cell receptor γδ cells. Compared with the ICH group, there were fewer apoptotic bodies and more Nissl bodies in the ICH + NSC group and the ICH + NSC + IL-17 group. To investigate the potential effect of IL-17 on directional differentiation of NSCs, we cultured mouse NSCs (NE-4C) alone or co-cultured them with T cell receptor γδ cells, which were isolated from mouse peripheral blood mononuclear cells, for 7 days. The results of western blot assays revealed that IL-17 secreted by T cell receptor γδ cells reduced the differentiation of NSCs into astrocytes and neurons, while IL-17 neutralization relieved the inhibition of directional differentiation into astrocytes rather than neurons. In conclusion, serum IL-17 levels were elevated in the early stage of ICH and were negatively correlated with outcome in ICH patients. Animal experiments and cytological investigations therefore demonstrated that IL-17 probably has neurotoxic roles in ICH because of its inhibitory effects on the directional differentiation of NSCs. The application of IL-17 neutralizing antibody may promote the directional differentiation of NSCs into astrocytes. This study was approved by the Clinical Research Ethics Committee of Anhui Medical University of China (For human study: Approval No. 20170135) in December 2016. All animal handling and experimentation were reviewed and approved by the Institutional Animal Care and Use Committee of Anhui Medical University (approval No. 20180248) in December 2017.
机译：白细胞介素17（IL-17）及其主要生产者T细胞受体γδ细胞，对脑出血（ICH）的发病机制具有神经毒性作用，加重脑损伤。为了研究IL-17和ICH之间的相关性，我们使用酶联免疫吸附测定动态筛选血清IL-17浓度，并探讨了ICH患者IL-17的临床值。在ICH患者中血清IL-17水平和神经恢复地位之间存在显着的负相关（R = -0.498，P <0.01）。为了研究IL-17的神经毒性作用，使用C57BL / 6小鼠来通过将自体血液注入尾部来建立ICH模型。随后，用小鼠神经干细胞（NSC）和/或IL-17中和抗体治疗小鼠72小时。流式细胞术，脑含水含量检测，NISL染色和末端脱氧核苷酸转移酶介导的DUTP缺口末端标记结果表明，NSC移植显着降低了PERI-血肿组织中的IL-17表达，但T细胞受体γδ细胞没有差异。与ICH组相比，ICH + NSC组中的细胞凋亡体和更多的尼斯菌体和ICH + NSC + IL-17组。为了探讨IL-17对NSCs的定向分化的潜在影响，我们单独培养小鼠NSCs（NE-4C）或用T细胞受体γδ细胞共培养它们，其与小鼠外周血单核细胞分离7天。 Western印迹测定结果显示，由T细胞受体γδ细胞分泌的IL-17将NSCs分化为星形胶质细胞和神经元的分化，而IL-17中和抑制了方向分化成星形胶质细胞而不是神经元。总之，血清IL-17水平在ICH的早期升高，并与ICH患者的结果负相关。因此，动物实验和细胞学调查表明IL-17可能在ICH中具有神经毒性作用，因为其对NSCs的定向分化的抑制作用。 IL-17中和抗体的应用可以促进NSCs进入星形胶质细胞的定向分化。 2016年12月，中国安徽医科大学临床研究伦理委员会批准了这项研究批准（针对人类学习：20170135号批准号2010135）。通过安徽医疗机构动物护理和使用委员会审查并批准了所有动物处理和实验大学（2018年12月批准No.201888）。

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