干扰素 γ 介导的 hsa-miR-424-5p 和 hsa-miR-513c-3p 失调与干燥综合征患者唾液腺功能障碍有关。
hsa-miR-424-5p and hsa-miR-513c-3p dysregulation mediated by IFN-γ is associated with salivary gland dysfunction in Sjögren's syndrome patients.
机构信息
Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Independencia 1027, 8380453, Independencia, Santiago, Chile.
Clínica INDISA, Av. Sta. María 1810, 7520440, Providencia, Santiago, Chile.
出版信息
J Autoimmun. 2023 Jul;138:103037. doi: 10.1016/j.jaut.2023.103037. Epub 2023 May 23.
Salivary secretory dysfunction in SS-patients is associated with altered proteostasis, upregulation of ATF6α and components of the ERAD complex, such as SEL1L, and downregulation of XBP-1s and GRP78. Hsa-miR-424-5p is downregulated and hsa-miR-513c-3p is overexpressed in salivary glands from SS-patients. These miRNAs emerged as candidates that could regulate ATF6/SEL1L and XBP-1s/GRP78 levels, respectively. This study aimed to evaluate the effect of IFN-γ on hsa-miR-424-5p and hsa-miR-513c-3p expression and how these miRNAs regulate their targets. In labial salivary glands (LSG) biopsies from 9 SS-patients and 7 control subjects and IFN-γ-stimulated 3D-acini were analyzed. hsa-miR-424-5p and hsa-miR-513c-3p levels were measured by TaqMan assays and their localization by ISH. mRNA, protein levels, and localization of ATF6, SEL1L, HERP, XBP-1s and GRP78 were determined by qPCR, Western blot, or immunofluorescence. Functional and interaction assays were also performed. In LSGs from SS-patients and IFN-γ-stimulated 3D-acini, hsa-miR-424-5p was downregulated and ATF6α and SEL1L were upregulated. ATF6α and SEL1L were decreased after hsa-miR-424-5p overexpression, while ATF6α, SEL1L and HERP increased after hsa-miR-424-5p silencing. Interaction assays revealed that hsa-miR-424-5p targets ATF6α directly. hsa-miR-513c-3p was upregulated and XBP-1s and GRP78 were downregulated. XBP-1s and GRP78 were decreased after hsa-miR-513c-3p overexpression, while increases in XBP-1s and GRP78 were observed after hsa-miR-513c-3p silencing. Furthermore, we determined that hsa-miR-513c-3p targets XBP-1s directly. Significant correlations were found between both miRNA levels and clinical parameters. In conclusion, IFN-γ-dependent hsa-miR-424-5p and hsa-miR-513c-3p levels affect the expression of important factors involved in cellular proteostasis that control secretory function in LSG from SS-patients.
干燥综合征患者的唾液分泌功能障碍与蛋白质稳态改变、ATF6α 上调和内质网相关降解复合物（ERAD）成分（如 SEL1L）上调以及 XBP-1s 和 GRP78 下调有关。干燥综合征患者的唾液腺中 hsa-miR-424-5p 下调，hsa-miR-513c-3p 表达上调。这些 miRNA 分别被认为可以调节 ATF6/SEL1L 和 XBP-1s/GRP78 的水平。本研究旨在评估 IFN-γ 对 hsa-miR-424-5p 和 hsa-miR-513c-3p 表达的影响，以及这些 miRNA 如何调节其靶标。我们分析了 9 名干燥综合征患者和 7 名对照受试者的唇腺（LSG）活检组织和 IFN-γ 刺激的 3D 腺泡，并通过 TaqMan 检测法测量 hsa-miR-424-5p 和 hsa-miR-513c-3p 的水平，通过原位杂交法检测其定位。通过 qPCR、Western blot 或免疫荧光法测定 ATF6、SEL1L、HERP、XBP-1s 和 GRP78 的 mRNA、蛋白水平和定位。还进行了功能和相互作用分析。在干燥综合征患者的 LSG 和 IFN-γ 刺激的 3D 腺泡中，hsa-miR-424-5p 下调，ATF6α 和 SEL1L 上调。hsa-miR-424-5p 过表达后，ATF6α 和 SEL1L 减少，而 hsa-miR-424-5p 沉默后，ATF6α、SEL1L 和 HERP 增加。相互作用分析表明 hsa-miR-424-5p 直接靶向 ATF6α。hsa-miR-513c-3p 上调，XBP-1s 和 GRP78 下调。hsa-miR-513c-3p 过表达后，XBP-1s 和 GRP78 减少，而 hsa-miR-513c-3p 沉默后，XBP-1s 和 GRP78 增加。此外，我们确定 hsa-miR-513c-3p 直接靶向 XBP-1s。两种 miRNA 水平与临床参数之间存在显著相关性。总之，IFN-γ 依赖性 hsa-miR-424-5p 和 hsa-miR-513c-3p 水平影响参与细胞蛋白质稳态的重要因素的表达，这些因素控制干燥综合征患者 LSG 的分泌功能。
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