Department of Neurology, the 2nd Affiliated Hospital of Harbin Medical University, Harbin, China.
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6881-6892. doi: 10.26355/eurrev_202006_21678.
T helper 17 (Th17) cells are involved in the pathogenesis of multiple sclerosis (MS). The present study aimed to explore the role of miR-1-3p in Th17 cell differentiation associated with MS. PATIENTS AND METHODS: Expression of miR-1-3p in periphery blood mononuclear cells (PBMC), cerebrospinal fluid (CSF), CD4+ T cells, CD8+ T cells, non-T cells and differentiated CD4+ T cells derived from healthy donors and MS patients during remitted and relapsed stages was detected. Level of ETS1 in PBMC in MS-relapse patients was examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Correlations among neurofilament light (NF-L), C-reactive protein (CRP), estrogen receptor 1 (ESR), interleukin 17A (IL-17A) in serum, NF-L, C-X-C motif chemokine ligand 13 (CXCL13), chitinase 3 like 1 (CHI3L1), RAR related orphan receptor C (RORC) in CSF, and ETS proto-oncogene 1 (ETS1), RORC in PBMC and miR-1-3p expression were analyzed. The target gene of miR-1-3p was analyzed by performing Dual-Luciferase reporter assay, and the IL-17A+ CD4+ (Th17) cells were detected by flow cytometer. Gene expressions of IL-17A, RORC and Th17 pathogenic were determined by qRT-PCR.
Upregulated miR-1-3p was observed in MS-relapse patients and Th17 cells, and expression of miR-1-3p was positively correlated with MS severity. MiR-1-3p overexpression in naïve CD4+ T cells promoted the differentiation of Th17 cells by upregulating the level of inflammation-associated markers. The expression of ETS1, which was predicted to be the target gene of miR-1-3p, was reduced in PBMC from MS-relapse patients, while the upregulation of ETS1 inhibited the expression of pathogenic genes of Th17.
The study demonstrated the positive role of miR-1-3p in Th17 differentiation associated with MS via targeting ETS1.
辅助性 T 细胞 17（Th17）细胞参与多发性硬化症（MS）的发病机制。本研究旨在探讨 miR-1-3p 在与 MS 相关的 Th17 细胞分化中的作用。
检测缓解期和复发期健康供者和 MS 患者外周血单个核细胞（PBMC）、脑脊液（CSF）、CD4+T 细胞、CD8+T 细胞、非 T 细胞和分化的 CD4+T 细胞中 miR-1-3p 的表达。采用定量实时聚合酶链反应（qRT-PCR）检测 MS 复发患者 PBMC 中的 ETS1 水平。分析血清中神经丝轻链（NF-L）、C 反应蛋白（CRP）、雌激素受体 1（ESR）、白细胞介素 17A（IL-17A）、CSF 中 NF-L、C-X-C 基序趋化因子配体 13（CXCL13）、几丁质酶 3 样 1（CHI3L1）、RAR 相关孤儿受体 C（RORC）与 PBMC 中的 ETS 原癌基因 1（ETS1）、RORC 和 miR-1-3p 表达之间的相关性。通过双荧光素酶报告基因实验分析 miR-1-3p 的靶基因，通过流式细胞术检测 IL-17A+CD4+（Th17）细胞。采用 qRT-PCR 检测 IL-17A、RORC 和 Th17 致病性基因的表达。
MS 复发患者和 Th17 细胞中观察到上调的 miR-1-3p，miR-1-3p 的表达与 MS 严重程度呈正相关。在幼稚 CD4+T 细胞中过表达 miR-1-3p 通过上调炎症相关标志物的水平促进 Th17 细胞的分化。预测为 miR-1-3p 靶基因的 ETS1 在 MS 复发患者 PBMC 中的表达减少，而 ETS1 的上调抑制了 Th17 致病性基因的表达。
本研究通过靶向 ETS1 证实了 miR-1-3p 在与 MS 相关的 Th17 分化中的积极作用。
