Department of Rheumatology, the Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu, China.
Department of Rheumatology, the Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
Biomed Pharmacother. 2019 Oct;118:109313. doi: 10.1016/j.biopha.2019.109313. Epub 2019 Aug 14.
Bone marrow-derived mesenchymal stem cells (MSCs) from systemic lupus erythematosus patients (SLE-BMSCs) exhibited abnormalities in cytokine production and immune modulation. Deregulation of Nod-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome plays an important role in SLE. Herein, we explored whether miRNAs are involved in the regulation of NLRP3 in SLE-BMSCs. ELISA assay was used to detect the levels of inflammatory cytokines. The expression levels of let-7f-5p and gene mRNAs were determined by qRT-PCR assay. The protein levels of NLRP3, Cleaved caspase-1 and ASC were measured by western blot. The interaction between let-7f-5p and NLRP3 was verified using dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. In vivo assay was performed to explore whether let-7f-5p upregulation could ameliorate inflammation in MRL/lpr mice. Our data indicated that SLE patients had significantly serum higher levels of IFN-γ, IL-6, IL-18, IL-12, IL-13 and IL-1β. We demonstrated that NLRP3 expression was upregulated in SLE-BMSCs. Let-7f-5p directly targeted NLRP3 and repressed NLRP3 expression. NLRP3 depletion or let-7f-5p upregulation repressed IL-1β production and the expression of NLRP3 inflammasome components. Moreover, upregulated let-7f-5p-mediated anti-inflammation effect was significantly abrogated by NLRP3 expression restoration. Besides, let-7f-5p upregulation ameliorated inflammation through modulating NLRP3 in vivo. In conclusion, our study suggested that high level of let-7f-5p alleviated inflammation in SLE-BMSCs at least partly through targeting NLRP3, highlighting let-7f-5p as a novel promising therapeutic strategy for SLE treatment.
骨髓间充质干细胞（MSCs）来源于系统性红斑狼疮患者（SLE-BMSCs），其细胞因子产生和免疫调节存在异常。核苷酸结合寡聚化结构域样受体热蛋白结构域包含蛋白 3（NLRP3）炎症小体的失调在系统性红斑狼疮中起着重要作用。在此，我们探讨了 microRNA 是否参与了 SLE-BMSCs 中 NLRP3 的调控。通过 ELISA 检测细胞因子水平。通过 qRT-PCR 检测 let-7f-5p 和基因 mRNAs 的表达水平。通过 Western blot 检测 NLRP3、Cleaved caspase-1 和 ASC 的蛋白水平。通过双荧光素酶报告基因实验和 RNA 免疫沉淀（RIP）实验验证 let-7f-5p 与 NLRP3 之间的相互作用。通过体内实验探讨 let-7f-5p 的上调是否可以改善 MRL/lpr 小鼠的炎症。我们的数据表明，系统性红斑狼疮患者的血清 IFN-γ、IL-6、IL-18、IL-12、IL-13 和 IL-1β 水平显著升高。我们证明 NLRP3 在 SLE-BMSCs 中表达上调。let-7f-5p 直接靶向 NLRP3 并抑制 NLRP3 的表达。NLRP3 耗竭或 let-7f-5p 的上调抑制了 IL-1β 的产生和 NLRP3 炎症小体组成部分的表达。此外，NLRP3 表达恢复显著削弱了 let-7f-5p 上调介导的抗炎作用。此外，let-7f-5p 上调通过体内调节 NLRP3 改善炎症。总之，我们的研究表明，高水平的 let-7f-5p 通过靶向 NLRP3 至少部分缓解了 SLE-BMSCs 的炎症，突显了 let-7f-5p 作为治疗系统性红斑狼疮的一种新的有前途的治疗策略。
