Arsenic trioxide ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice by inducing CD4+ T cell apoptosis
Arsenic trioxide ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice by inducing CD4+ T cell apoptosis
复制标题
三氧化二砷通过诱导 CD4( ) T 细胞凋亡改善 C57BL/6 小鼠实验性自身免疫性脑脊髓炎
DOI：
10.1186/s12974-020-01829-x
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发表时间：
2020-05-06
影响因子：
9.3
通讯作者：
Xia, Jun-Jie
中科院分区：
文献类型：
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作者：
An, Ke;Xue, Meng-Jiao;Xia, Jun-Jie
背景 多发性硬化症（MS）是一种中枢神经系统的免疫介导性疾病，其特征为严重的白质脱髓鞘。由于其发病机制复杂，目前尚无根治MS的方法。实验性自身免疫性脑脊髓炎（EAE）是研究MS的理想动物模型。三氧化二砷（ATO）是一种具有治疗多种自身免疫性疾病功效的中药。因其具有抗炎和免疫抑制特性，它也被用于抑制急性免疫排斥反应。然而，ATO对EAE是否具有治疗作用尚不清楚，其潜在机制也尚未明确阐明。在本研究中，我们试图评估ATO是否可用于改善小鼠的EAE。
方法 在免疫接种10天后，对EAE小鼠腹腔注射ATO（0.5毫克/千克/天），持续8天。在免疫接种第22天，采集脊髓、脾脏和血液，以分析脱髓鞘情况、炎症反应、小胶质细胞活化情况以及CD4(+) T细胞的比例。在体外，为进行机制研究，从健康C57BL/6小鼠的脾脏中分离出CD4(+) T细胞，用ATO处理，然后用于细胞凋亡检测、JC - 1染色、透射电子显微镜成像以及蛋白质免疫印迹分析。
结果 ATO延缓了EAE的发病，并减轻了小鼠EAE的严重程度。ATO处理还减轻了脱髓鞘现象，缓解了炎症，减少了小胶质细胞的活化，并降低了EAE小鼠体内白细胞介素 - 2（IL - 2）、干扰素 - γ（IFN - γ）、白细胞介素 - 1β（IL - 1β）、白细胞介素 - 6（IL - 6）和肿瘤坏死因子 - α（TNF - α）的表达水平。此外，在接受ATO治疗的EAE小鼠中，脊髓、脾脏和外周血中CD4(+) T细胞的数量和比例均有所减少。最后，ATO在体外和体内均通过线粒体途径诱导CD4(+) T细胞凋亡。另外，给予ATO对心脏、肝脏或肾脏功能没有不良影响，也不会诱导脊髓细胞凋亡。
结论 总体而言，我们的研究结果表明，ATO在EAE的起始和进展过程中发挥保护作用，有潜力成为治疗MS的一种新型药物。
Background Multiple sclerosis (MS) is an immune-mediated disease of the central nervous system characterized by severe white matter demyelination. Because of its complex pathogenesis, there is no definite cure for MS. Experimental autoimmune encephalomyelitis (EAE) is an ideal animal model for the study of MS. Arsenic trioxide (ATO) is an ancient Chinese medicine used for its therapeutic properties with several autoimmune diseases. It is also used to inhibit acute immune rejection due to its anti-inflammatory and immunosuppressive properties. However, it is unclear whether ATO has a therapeutic effect on EAE, and the underlying mechanisms have not yet been clearly elucidated. In this study, we attempted to assess whether ATO could be used to ameliorate EAE in mice. Methods ATO (0.5 mg/kg/day) was administered intraperitoneally to EAE mice 10 days post-immunization for 8 days. On day 22 post-immunization, the spinal cord, spleen, and blood were collected to analyze demyelination, inflammation, microglia activation, and the proportion of CD4(+) T cells. In vitro, for mechanistic studies, CD4(+) T cells were sorted from the spleen of naive C57BL/6 mice and treated with ATO and then used for an apoptosis assay, JC-1 staining, imaging under a transmission electron microscope, and western blotting. Results ATO delayed the onset of EAE and alleviated the severity of EAE in mice. Treatment with ATO also attenuated demyelination, alleviated inflammation, reduced microglia activation, and decreased the expression levels of IL-2, IFN-gamma, IL-1 beta, IL-6, and TNF-alpha in EAE mice. Moreover, the number and proportion of CD4(+) T cells in the spinal cord, spleen, and peripheral blood were reduced in ATO-treated EAE mice. Finally, ATO induced CD4(+) T cell apoptosis via the mitochondrial pathway both in vitro and in vivo. Additionally, the administration of ATO had no adverse effect on the heart, liver, or kidney function, nor did it induce apoptosis in the spinal cord. Conclusions Overall, our findings indicated that ATO plays a protective role in the initiation and progression of EAE and has the potential to be a novel drug in the treatment of MS.