Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China.
Mediators Inflamm. 2020 Sep 30;2020:4286047. doi: 10.1155/2020/4286047. eCollection 2020.
This study is aimed at exploring the expression pattern and methylation level of in the peripheral blood mononuclear cells (PBMCs) of myasthenia gravis (MG) patients with positive acetylcholine receptor (AChR) autoantibodies and revealing the relationship between the methylation pattern and MG. The relationship between the NFAT family members and was explored to reveal the regulatory mechanism of in the pathogenesis and treatment of AChR MG. Moreover, we attempted to demonstrate the potential therapeutic mechanism of tacrolimus in AChR MG. The relative expression level in the PBMCs of healthy people was compared with that in the PBMCs of AChR MG patients with quantitative real-time PCR (qRT-PCR). The methylation frequency of the promoter was detected by bisulfite sequencing PCR (BSP) and pyrosequencing. A dual-luciferase reporter system was used to reveal the relationship between the promoter and nuclear factor of activated T cells 5 (). The qRT-PCR results showed that expression was significantly upregulated in the B cells and CD8+ T cells of AChR MG patients but not in the CD4+ T cells, and these expression differences were significantly associated with a decrease in methylation. , which was speculated to bind to island 1 (p1) in the promoter, may regulate the lymphocyte balance by regulating gene expression but failed to affect the methylation of the promoter. Tacrolimus therapy-induced methylation and overexpression of could significantly reduce the expression of in AChR MG patients. The gene was remarkably upregulated in the PBMCs of MG patients. may affect transcription initiation and downregulate expression through p1 in the promoter, thus controlling gene expression and regulating the lymphocyte balance. Therefore, could be an immune regulatory factor in both AChR MG occurrence and treatment with tacrolimus.
本研究旨在探讨乙酰胆碱受体（AChR）自身抗体阳性的重症肌无力（MG）患者外周血单个核细胞（PBMC）中 的表达模式和甲基化水平，并揭示 甲基化模式与 MG 之间的关系。通过探索 NFAT 家族成员与 的关系，揭示 在 AChR MG 发病机制和治疗中的调控机制。此外，我们试图证明他克莫司在 AChR MG 中的潜在治疗机制。通过实时定量 PCR（qRT-PCR）比较健康人和 AChR MG 患者 PBMC 中 的相对表达水平。通过亚硫酸氢盐测序 PCR（BSP）和焦磷酸测序检测 的启动子甲基化频率。双荧光素酶报告系统揭示 启动子与活化 T 细胞核因子 5（NFAT5）之间的关系。qRT-PCR 结果显示，AChR MG 患者 B 细胞和 CD8+T 细胞中 的表达显著上调，而 CD4+T 细胞中则没有上调，这些表达差异与 甲基化减少显著相关。推测 可能通过调节 基因表达来调节淋巴细胞平衡，但未能影响 的启动子甲基化。他克莫司治疗诱导的 甲基化和过表达可显著降低 AChR MG 患者 的表达。 在 MG 患者的 PBMC 中显著上调。 可能通过启动子中的 p1 影响转录起始并下调 的表达，从而控制 的基因表达并调节淋巴细胞平衡。因此， 可能是 AChR MG 发病和他克莫司治疗中免疫调节因子。
