a Department of Rheumatology and Immunology, The Affiliated Hospital of Qingdao University , Shandong , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2473-2480. doi: 10.1080/21691401.2019.1626413.
The interaction of long non-coding RNAs (lncRNAs)-microRNAs (miRs) exerts crucial functions in mediating inflammatory reaction. It is still unclear whether myocardial infarction associated transcript 2 (Mirt2)-miR-377 mediates the inflammatory pathogenesis in Sjögren's syndrome (SS). The inflammatory lesion model was established by stimulating salivary gland epithelial cells (SGECs) by interferon gamma (IFN-γ). Mirt2- and/or miR-377-transfected SGECs, as well as their negative controls, were applied to investigate the biological functions in inflammation. Cell viability and apoptosis were examined using commercial kits. Western blot was applied to quantify protein level, and enzyme-linked immuno sorbent assay (ELISA) was used to value the secretion of cytokines. The up-regulation of Mirt2 was observed in IFN-γ-treated SGECs. Mirt2 overexpression restored the expression of miR-377 which was repressed by IFN-γ. However, miR-377 silence abolished the protective effect on cell viability, inhibitory effect on apoptosis and prohibitive role in pro-inflammatory factors. Mirt2 diminished the phosphorylated expression of crucial regulators while miR-377 silence restored the phosphorylation in IFN-γ-treated SGECs. Mirt2 was elevated in IFN-γ-treated SGECs and then up-regulated miR-377 in response to inflammatory lesions. Mechanically, in synergy with miR-377 Mirt2 blocked IFN-γ-evoked activation of NF-κB and JAK/STAT signalling pathway.
长链非编码 RNA（lncRNA）-microRNA（miR）的相互作用在介导炎症反应中发挥着重要作用。心肌梗塞相关转录物 2（Mirt2）-miR-377 是否介导干燥综合征（SS）中的炎症发病机制尚不清楚。通过干扰素 γ（IFN-γ）刺激唾液腺上皮细胞（SGEC）建立炎症损伤模型。将 Mirt2-和/或 miR-377 转染的 SGEC 及其阴性对照应用于炎症中的生物学功能研究。使用商业试剂盒检测细胞活力和细胞凋亡。Western blot 用于定量蛋白水平，酶联免疫吸附测定（ELISA）用于评估细胞因子的分泌。IFN-γ 处理的 SGEC 中观察到 Mirt2 的上调。Mirt2 过表达恢复了 miR-377 的表达，miR-377 被 IFN-γ 抑制。然而，miR-377 沉默消除了对细胞活力的保护作用、对细胞凋亡的抑制作用以及对促炎因子的抑制作用。Mirt2 降低了关键调节因子的磷酸化表达，而 miR-377 沉默恢复了 IFN-γ 处理的 SGEC 中的磷酸化。IFN-γ 处理的 SGEC 中 Mirt2 升高，然后响应炎症损伤而上调 miR-377。在机制上，Mirt2 与 miR-377 协同作用阻断了 IFN-γ 诱导的 NF-κB 和 JAK/STAT 信号通路的激活。
