Transcriptional profiling of murine macrophages stimulated with cartilage fragments revealed a strategy for treatment of progressive osteoarthritis
Transcriptional profiling of murine macrophages stimulated with cartilage fragments revealed a strategy for treatment of progressive osteoarthritis
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DOI：
10.1038/s41598-020-64515-1
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发表时间：
2020-05-05
影响因子：
4.6
通讯作者：
Iwasaki, Norimasa
中科院分区：
文献类型：
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作者：
Hamasaki, Masanari;Terkawi, Mohamad Alaa;Iwasaki, Norimasa
越来越多的证据表明，滑膜炎与骨关节炎过程有关。巨噬细胞在滑膜炎的发生发展中起主要作用。我们早期的研究表明，软骨碎片和巨噬细胞之间的相互作用加剧了骨关节炎的过程。然而，软骨碎片触发细胞反应的分子机制仍有待研究。因此，本研究旨在分析巨噬细胞对软骨碎片的分子反应。为此，我们通过RNA测序分析了暴露于软骨碎片的小鼠巨噬细胞的转录图谱。共有153个基因因软骨碎片而表达上调，105个基因表达下调。生物信息学分析表明，上调基因中最显著丰富的术语包括清道夫受体活性、整合素结合活性、肿瘤坏死因子信号转导和Toll样受体信号转导。为了进一步证实我们的结果，我们对OA患者滑膜组织中的调节分子进行了免疫组织化学染色。与RNA-SEQ结果一致，Marco、TLR2和ITGα5主要表达于滑膜组织的内膜衬里层。此外，阻断TLR2或ITGα5，但不使用MARCO特异性抗体，可显著减少软骨碎片刺激的巨噬细胞产生的肿瘤坏死因子-α。我们的数据表明，阻断TLR2或ITGα5可能是治疗进行性骨关节炎的有前途的治疗策略。
Accumulating evidence suggests that synovitis is associated with osteoarthritic process. Macrophages play principal role in development of synovitis. Our earlier study suggests that interaction between cartilage fragments and macrophages exacerbates osteoarthritic process. However, molecular mechanisms by which cartilage fragments trigger cellular responses remain to be investigated. Therefore, the current study aims at analyzing molecular response of macrophages to cartilage fragments. To this end, we analyzed the transcriptional profiling of murine macrophages exposed to cartilage fragments by RNA sequencing. A total 153 genes were differentially upregulated, and 105 genes were down-regulated in response to cartilage fragments. Bioinformatic analysis revealed that the most significantly enriched terms of the upregulated genes included scavenger receptor activity, integrin binding activity, TNF signaling, and toll-like receptor signaling. To further confirm our results, immunohistochemical staining was performed to detected regulated molecules in synovial tissues of OA patients. In consistence with RNA-seq results, MARCO, TLR2 and ITG alpha 5 were mainly detected in the intima lining layer of synovial tissues. Moreover, blockade of TLR2 or ITG alpha 5 but not Marco using specific antibody significantly reduced production of TNF-alpha in stimulated macrophages by cartilage fragments. Our data suggested that blocking TLR2 or ITG alpha 5 might be promising therapeutic strategy for treating progressive osteoarthritis.