IL-17RA aptamer-mediated repression of IL-6 inhibits synovium inflammation in a murine model of osteoarthritis
IL-17RA aptamer-mediated repression of IL-6 inhibits synovium inflammation in a murine model of osteoarthritis
复制标题
IL-17RA 适体介导的 IL-6 抑制抑制小鼠骨关节炎模型中的滑膜炎症
DOI：
10.1016/j.joca.2011.01.018
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发表时间：
2011-06-01
影响因子：
7
通讯作者：
Liu, S. Q.
中科院分区：
文献类型：
--
作者：
Chen, L.;Li, D. Q.;Liu, S. Q.
目的：方法：采用新的细胞-SELEX技术，获得特异性的IL-17 RA核酸适配子，并将其应用于实验性骨关节炎（OA）小鼠模型中。合成了具有四（30）个探针的单链（ss）DNA库。通过将该文库与NIH 3 T3细胞孵育，我们收集了可以结合细胞表面的DNA配体。将收集的配体与IL-17 RA缺陷型NIH 3 T3细胞一起孵育，并从上清液中收获未结合的ssDNA用于下一轮选择。12个循环后，产生了针对IL-17 RA的特异性适体。对于动物实验，在Balb/C小鼠上进行椎间盘切除术以产生OA的动物模型。小鼠每周接受关节内（i.a.）注射适体或对照治疗6周。结果：筛选到一种能有效阻断IL-17与IL-17 RA结合的适配体RA 10 -6，并呈剂量依赖性。组织学检查和定量RT-PCR结果显示，RA 10 -6，特别是与塞来昔布联合注射的OA小鼠表现出抑制滑膜增厚和降低滑膜组织中IL-6的水平。结论：我们的结果表明，RA 10 -6可以抑制滑膜炎症通过阻断IL-17/1 L-17 RA介导的IL-6的表达。RA 10 -6与塞来昔布协同抑制滑膜组织中IL-6的表达。因此，靶向IL-17 RA的适体可能作为早期治疗OA的有效促炎剂。(C)2011年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Generate DNA aptamers to inhibit IL-17RA-mediated synovial inflammation in an experimental mouse model of osteoarthritis (OA).Methods: A novel cell-SELEX method was applied to obtain DNA aptamers specific for IL-17RA. A single-stranded (ss) DNA library with four(30) probes was synthesised. By incubating this library with NIH3T3 cells, we collected DNA ligands that could bind the cell surface. The collected ligands were incubated with IL-17RA-deficient NIH3T3 cells, and unbound ssDNA was harvested from the supernatant for the next round of selection. After 12 cycles, specific aptamers against IL-17RA were generated. For animal experiments, a meniscectomy was performed on Balb/C mice to generate an animal model of OA. Mice received weekly intra-articular (i.a.) injections of aptamers or control treatments for 6 weeks. Synovial membranes were evaluated by histomorphology and the mRNAs of critical inflammatory cytokines were measured by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).Results: An aptamer termed RA10-6 was obtained that could efficiently block IL-17 binding to IL-17RA in a dose-dependent manner in vitro. Histological examination and quantitative RT-PCR results showed that OA mice that injected with RA10-6, especially in combination with celecoxib demonstrated inhibition of synovial thickening and reduction in IL-6 levels in the synovial tissue.Conclusion: Our results suggest that RA10-6 can inhibit synovial inflammation by blocking IL-17/1L-17RA-mediated IL-6 expression. RA10-6 acted synergistically with celecoxib to inhibit IL-6 expression in synovial tissues. Thus, aptamers targeting IL-17RA might serve as potent adjunctive agents for the early treatment of OA. (C) 2011 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.