Let-7f-5p ameliorates inflammation by targeting NLRP3 in bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus
Let-7f-5p ameliorates inflammation by targeting NLRP3 in bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus
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Let-7f-5p 通过靶向系统性红斑狼疮患者骨髓间充质干细胞中的 NLRP3 改善炎症
DOI：
10.1016/j.biopha.2019.109313
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发表时间：
2019-10-01
影响因子：
7.5
通讯作者：
Li, Guoqing
中科院分区：
文献类型：
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作者：
Tan, Wei;Gu, Zhifeng;Li, Guoqing
系统性红斑狼疮患者骨髓间充质干细胞（SLE-BMSCs）在细胞因子产生和免疫调节方面表现出异常。Nod样受体pyrin domain containing protein 3（NLRP 3）炎性小体的失调在SLE中起重要作用。在此，我们探讨了miRNA是否参与了SLE BMSCs中NLRP 3的调控。采用ELISA法检测炎症细胞因子水平。通过qRTPCR测定来确定let-7 f-5 p和基因mRNA的表达水平。Western blot检测NLRP 3、Cleaved caspase-1和ASC蛋白水平。let-7 f-5 p和NLRP 3之间的相互作用使用双荧光素酶报告基因测定和RNA免疫沉淀（RIP）测定来验证。进行体内测定以探索let-7 f-5 p上调是否可以改善MRL/lpr小鼠中的炎症。我们的数据表明，SLE患者血清中IFN-γ、IL-6、IL-18、IL-12、IL-13和IL-1 β水平显著升高。我们证明了NLRP 3在SLE-BMSCs中的表达上调。Let-7 f-5 p直接靶向NLRP 3并抑制NLRP 3表达。NLRP 3耗竭或let-7 f-5 p上调抑制IL-1 β产生和NLRP 3炎性体组分的表达。此外，上调的let-7 f-5 p介导的抗炎作用被NLRP 3表达恢复显著消除。此外，let-7 f-5 p上调通过调节体内NLRP 3而减轻炎症。总之，我们的研究表明，高水平的let-7 f-5 p至少部分地通过靶向NLRP 3减轻了SLE-BMSC中的炎症，突出了let-7 f-5 p作为SLE治疗的新的有希望的治疗策略。
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 qRTPCR 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-gamma, IL-6, IL-18, IL-12, IL-13 and IL-1 beta. 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-1 beta 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.