The upregulation of miR-98-5p affects the glycosylation of IgA1 through cytokines in IgA nephropathy
The upregulation of miR-98-5p affects the glycosylation of IgA1 through cytokines in IgA nephropathy
复制标题
IgA肾病中miR-98-5p的上调通过细胞因子影响IgA1的糖基化
DOI：
10.1016/j.intimp.2020.106362
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发表时间：
2020-05-01
影响因子：
5.6
通讯作者：
Liu, Hong
中科院分区：
文献类型：
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作者：
Liu, Di;Xia, Ming;Liu, Hong
目的：半乳糖缺陷型 IgA1（Gd-IgA1）的增加在 IgA 肾病（IgAN）的发病机制中发挥着至关重要的作用，最近的一些实验表明 microRNA（miRNA）参与调节肾脏的发育和生理功能。本研究的目的是鉴定影响IgAN发病机制的miRNA，揭示外周血中IgA1糖基化的潜在调控机制。方法：通过高通量测序筛选IgAN患者和健康对照者外周血单核细胞（PBMC）中差异表达的miRNA，并预测这些miRNA的靶点并进行分析。 通过双荧光素酶报告基因测定验证。我们还通过转染miRNA模拟物和相关质粒探讨了miRNA对Gd-IgA1的调控。结果：高通量测序结果显示，与健康对照相比，IgAN患者的PBMC中miR-98-5p的表达更高，荧光素酶报告基因系统证实miR-98-5p可能靶向趋化因子配体3（CCL3）。 si-CCL3的转染证实CCL3的减少可以影响白细胞介素6（IL-6）和C1GALT1的表达。通过转染miR-98-5p模拟物在PBMCs中过度表达miR-98-5p，降低了CCL3和C1GALT1水平并增加了IL-6水平，并且通过与CCL3质粒共转染来减弱PBMCs中的这些变化。 结论：结果表明，在PBMCs中，miR-98-5p可以靶向CCL3以降低其表达 从而增加 IL-6 水平，而由此产生的 IL-6 增加可以减少 C1GALT1 表达。因此，miR-98-5p可能参与IgAN的发生。
Objective: Increases in galactose-deficient IgA1 (Gd-IgA1) play a crucial role in the pathogenesis of IgA nephropathy (IgAN), and several recent experiments have shown that microRNAs (miRNAs) are involved in regulating the development and physiological function of the kidney. The aims of this study were to identify miRNAs that can affect the pathogenesis of IgAN and reveal the underlying regulatory mechanism of IgA1 glycosylation in peripheral blood.Methods: The differentially expressed miRNAs in peripheral blood mononuclear cells (PBMCs) between IgAN patients and healthy controls were screened by high-throughput sequencing, and the targets of these miRNAs were predicted and verified by dual-luciferase reporter assays. We also explored the miRNA regulation of Gd-IgA1 through the transfection of miRNA mimics and related plasmids.Results: The high-throughput sequencing results showed that miR-98-5p was more highly expressed in the PBMCs of IgAN patients compared with healthy controls, and the luciferase reporter gene system confirmed that miR-98-5p might target chemokine ligand 3 (CCL3). The transfection of si-CCL3 confirmed that a decrease in CCL3 can affect the expression of interleukin-6 (IL-6) and C1GALT1. The overexpression of miR-98-5p in PBMCs through the transfection of miR-98-5p mimic reduced the CCL3 and C1GALT1 levels and increased the IL-6 levels, and these changes in PBMCs were attenuated by cotransfection with the CCL3 plasmid.Conclusion: The results showed that in PBMCs, miR-98-5p can target CCL3 to decrease its expression and thereby increase the IL-6 levels, and the resulting increase in IL-6 can decrease C1GALT1 expression. Therefore, miR-98-5p might be involved in the development of IgAN.