Department of Metabolism and Endocrinology, The Second Xiangya Hospital, Central South University, Changsha, 410011, Hunan, China.
Key Laboratory of Diabetes Immunology (Central South University), Ministry of Education, National Clinical Research Center for Metabolic Diseases, Changsha, 410011, Hunan, China.
Acta Diabetol. 2021 Jan;58(1):63-72. doi: 10.1007/s00592-020-01591-9. Epub 2020 Aug 20.
Autoimmune diabetes mellitus (defined as ADM) comprises classical type 1 diabetes mellitus (T1DM) and latent autoimmune diabetes in adults (LADA). In this study, microRNAs (miRNAs) expression profiles and functions in peripheral blood mononuclear cells (PBMCs) of ADM patients were mapped and used to explore epigenetic regulation of the pathogenesis of ADM.
PBMCs samples from T1DM patients, LADA patients, and type 2 diabetes mellitus (T2DM) patients, as well as age- and sex-matched healthy controls for T1DM and T2DM, respectively, were collected and were sequenced to screen the miRNAs expression profiles. The target genes were verified by dual-luciferase reporter assay. Silencing or overexpressing of the differentially expressed miRNAs, or simultaneously silencing the miRNAs and it's target gene, and then levels of the mRNAs, protein and cytokines were detected.
miR-143-3p expression was upregulated in ADM patients. The target gene of miR-143-3p was identified as Fos-related antigen 2 (FOSL2). Transfection of a miR-143-3p inhibitor into PBMCs upregulated FOSL2 expression, resulting in a downregulated expression of the IL-2, TNF-α, and IFN-γ, and an upregulated expression of IL-6. Transfection of a miR-143-3p mimic into PBMCs downregulated FOSL2 expression, leading to an upregulation of IL-2 and TNF-α expression and a downregulation of IL-6 expression. When silencing FOSL2 while inhibiting miR-143-3p in PBMCs, there was no significant change in expression of the FOSL2 mRNA, protein and cytokines.
The expression of miR-143-3p in PBMCs from ADM patients is upregulated. miR-143-3p could function in the pathogenesis of ADM by modulating the inflammatory reaction through FOSL2.
自身免疫性糖尿病（定义为 ADM）包括经典 1 型糖尿病（T1DM）和成人隐匿性自身免疫性糖尿病（LADA）。在这项研究中，我们绘制了 ADM 患者外周血单个核细胞（PBMC）中的 microRNAs（miRNAs）表达谱和功能，并用于探索 ADM 发病机制的表观遗传调控。
收集 T1DM 患者、LADA 患者和 2 型糖尿病（T2DM）患者的 PBMCs 样本，以及与 T1DM 和 T2DM 年龄和性别匹配的健康对照者的 PBMCs 样本，进行测序以筛选 miRNAs 表达谱。通过双荧光素酶报告基因实验验证靶基因。然后通过沉默或过表达差异表达的 miRNAs，或同时沉默 miRNAs 和其靶基因，并检测 mRNAs、蛋白质和细胞因子的水平。
ADM 患者的 miR-143-3p 表达上调。miR-143-3p 的靶基因被鉴定为 Fos 相关抗原 2（FOSL2）。将 miR-143-3p 抑制剂转染至 PBMCs 中可上调 FOSL2 表达，导致 IL-2、TNF-α 和 IFN-γ 的表达下调，IL-6 的表达上调。将 miR-143-3p 模拟物转染至 PBMCs 中可下调 FOSL2 表达，导致 IL-2 和 TNF-α 表达上调，IL-6 表达下调。当在 PBMCs 中沉默 FOSL2 同时抑制 miR-143-3p 时，FOSL2 mRNA、蛋白质和细胞因子的表达没有明显变化。
ADM 患者 PBMCs 中的 miR-143-3p 表达上调。miR-143-3p 可通过 FOSL2 调节炎症反应在 ADM 的发病机制中发挥作用。
