MicroRNA-20a-5p suppresses IL-17 production by targeting OSM and CCL1 in patients with Vogt-Koyanagi-Harada disease
MicroRNA-20a-5p suppresses IL-17 production by targeting OSM and CCL1 in patients with Vogt-Koyanagi-Harada disease
复制标题
MicroRNA-20a-5p 通过靶向 OSM 和 CCL1 抑制 Vogt-Koyanagi-Harada 病患者的 IL-17 产生
DOI：
10.1136/bjophthalmol-2017-311079
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发表时间：
2018-02-01
影响因子：
4.1
通讯作者：
Yang, Peizeng
中科院分区：
文献类型：
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作者：
Chang, Rui;Yi, Shenglan;Yang, Peizeng
目的探讨microRNA-20 a-5p（miR-20 a-5p）在Vogt-Koyanagi-Harada（VKH）病发病机制中的作用。亚硫酸氢盐测序PCR检测miR-20a-5p启动子甲基化状态。通过荧光素酶报告基因测定评价靶标。结果与正常对照组相比，活动性VKH患者外周血CD4（+）T细胞中miR-20 a-5p表达水平显著降低，而活动性VKH患者外周血CD4（+）T细胞中miR-20 a-5p表达水平显著降低。这两个基因，抑瘤素M（OSM）和C-C基序趋化因子配体1（CCL1），被确定为miR-20a-5p的靶基因。miR-20a-5p的上调显著抑制了活动性VKH患者CD4（+）T细胞中白细胞介素17（IL-17）的产生，而miR-20a-5p的下调则表现出相反的作用。此外，OSM和CCL1的过表达可以挽救miR-20a-5p上调的效果。此外，miR-20a-5p的水平响应于启动子的超甲基化而降低。结论miR-20a-5p的下调可能是由于启动子区甲基化所致。miR-20 a-5p还可以通过靶向活动性VKH患者CD4（+）T细胞中的OSM和CCL1产生来抑制IL-17的产生。
Aim To elucidate the role of microRNA-20a-5p (miR-20a-5p) in the pathogenesis of Vogt-Koyanagi-Harada (VKH) disease.Methods Quantitative real-time PCR was used to quantify miR-20a-5p expression in CD4(+) T cells from patients with active VKH and normal controls. The promoter methylation status of miR-20a-5p was detected by bisulfite sequencing PCR. Targets were evaluated by a luciferase reporter assay. The functional effects of miR-20a-5p on CD4(+) T cells from patients with active VKH were assessed by upregulation or downregulation of its expression using liposomes.Results The miR-20a-5p level was significantly decreased in CD4(+) T cells from patients with active VKH as compared with normal controls. The two genes, oncostatin M (OSM) and C-C motif chemokine ligand 1 (CCL1), were identified as targets of miR-20a-5p. The upregulation of miR-20a-5p significantly suppressed interleukin 17 (IL-17) production in CD4(+) T cells from patients with active VKH, whereas downregulation of miR-20a-5p exhibited an inverse effect. In addition, overexpression of OSM and CCL1 could rescue the effect of the upregulation of miR-20a-5p. Moreover, the level of miR-20a-5p was reduced in response to hypermethylation of the promoter. Further study showed that miR-20a-5p suppressed the activity of the phosphoinositide 3-kinase-AKT pathway.Conclusions Our findings indicate that downregulation of miR-20a-5p is caused by promoter hypermethylation. MiR-20a-5p could also suppress the production of IL-17 by targeting OSM and CCL1 production in CD4(+) T cells in patients with active VKH.