Downregulation of miR-145-5p contributes to hyperproliferation of keratinocytes and skin inflammation in psoriasis
Downregulation of miR-145-5p contributes to hyperproliferation of keratinocytes and skin inflammation in psoriasis
复制标题
miR-145-5p 的下调导致牛皮癣中角质形成细胞过度增殖和皮肤炎症
DOI：
10.1111/bjd.17256
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发表时间：
2019-02-01
影响因子：
10.3
通讯作者：
Sun, Q.
中科院分区：
文献类型：
--
作者：
Yan, J. J.;Qiao, M.;Sun, Q.
背景：微rna （miRNAs）在牛皮癣发病机制中的广泛参与已被充分证实。然而，关于特定mirna对这种疾病的患病率的贡献知之甚少。目的探讨miR-145-5p在银屑病中的作用。方法对4例银屑病患者和4例对照组进行miRNA芯片分析。采用定量逆转录酶聚合酶链反应和荧光原位杂交技术鉴定表达异常的mirna。采用荧光素酶测定来确定miR-145-5p是否靶向混合谱系激酶（MLK）3。CCK-8法和磁性荧光法检测细胞增殖和趋化因子分泌。Western blot检测MLK3及其下游效应物的蛋白表达水平。建立银屑病小鼠模型进行体内实验。结果miR-145-5p在银屑病皮损中表达下调。荧光素酶检测显示MLK3是miR-145-5p的直接靶点。正常人表皮角质形成细胞（NHEKs）中miR-145-5p的过表达抑制了细胞增殖和趋化因子的分泌。相反，沉默miR-145-5p可促进NHEK增殖并增加趋化因子分泌。沉默MLK3消除了miR-145-5p抑制剂诱导的细胞增殖和趋化因子表达的促进。miR-145-5p通过靶向MLK3调控核因子κ B和转录信号转导及激活因子3。向皮肤内递送agomiR-145-5p可减少表皮增生并改善牛皮癣样皮炎。给药antagomiR-145-5p则产生相反的效果。我们的研究结果表明，miR-145-5p通过靶向MLK3负向调节NHEKs的增殖和趋化因子的分泌，miR-145-5p的下调有助于银屑病皮损的皮肤炎症。
Background The extensive involvement of microRNAs (miRNAs) in the pathogenesis of psoriasis is well documented. However, little is known about the contribution of specific miRNAs to the prevalence of this disease. Objectives To explore the role of miR-145-5p in psoriasis. Methods miRNA microarray analysis was performed in four patients with psoriasis and four controls. Quantitative reverse-transcriptase polymerase chain reaction and fluorescence in situ hybridization were used to identify the dysregulated miRNAs. Luciferase assays were performed to determine whether miR-145-5p targets mixed-lineage kinase (MLK)3. CCK-8 assay and Magnetic Luminex Assay were performed to measure cell proliferation and chemokine secretion. Western blot analysis was used to investigate the protein levels of MLK3 and its downstream effectors. Mouse models of psoriasis were established for in vivo experiments. Results miR-145-5p was downregulated in psoriatic lesional skin. Luciferase assays showed that MLK3 is a direct target of miR-145-5p. Overexpression of miR-145-5p in normal human epidermal keratinocytes (NHEKs) suppressed cell proliferation and secretion of chemokines. In contrast, silencing miR-145-5p promoted NHEK proliferation and increased chemokine secretion. Silencing MLK3 abrogated miR-145-5p inhibitor-induced promotion of cell proliferation and chemokine expression. miR-145-5p regulates nuclear factor-kappa B and signal transducer and activator of transcription 3 by targeting MLK3. Delivery of agomiR-145-5p into the skin decreased epidermal hyperplasia and ameliorated psoriasis-like dermatitis. Delivery of antagomiR-145-5p led to the opposite effects. Conclusions Our findings indicate that miR-145-5p negatively regulates proliferation and chemokine secretion of NHEKs by targeting MLK3, and downregulation of miR-145-5p contributes to skin inflammation in psoriasis lesions.