miR-640 aggravates intervertebral disc degeneration via NF-κB and WNT signalling pathway
miR-640 aggravates intervertebral disc degeneration via NF-κB and WNT signalling pathway
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DOI：
10.1111/cpr.12664
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发表时间：
2019-07-25
影响因子：
8.5
通讯作者：
Qian, Jixian
中科院分区：
文献类型：
--
作者：
Dong, Wengang;Liu, Jun;Qian, Jixian
目的腰背痛是当今骨科的常见病。它主要由椎间盘退变引起。本研究试图揭示目前尚不清楚的退行性变的发病机制和治疗策略。材料与方法收集15例退变腰椎间盘组织和5例健康志愿者。对髓核和纤维环细胞进行传代培养。通过qPCR测定miR-640表达。计算机分析和荧光素酶报告基因分析用于确定miR-640靶基因。免疫组织化学和免疫细胞化学染色用于追踪促炎细胞因子和信号传导通路的关键转导子。我们还使用β-半乳糖苷酶染色、流式细胞术和细胞活力测定来监测退化指数。结果miR-640在退变髓核组织和细胞中过表达。炎症环境通过NF-κ B信号通路促进miR-640表达。此外，miR-640靶向LRP 1并增强NF-κ B信号活性，从而构建了正反馈环。miR-640抑制beta-catenin和EP 300的表达，从而抑制WNT信号，导致髓核细胞变性。miR-640抑制剂处理在体外表现出抗炎、逆转WNT信号传导途径耗竭和缓解退行性特征的作用。结论miR-640在椎间盘退变及相关炎症微环境中起重要作用。它是下腰痛生物治疗的一个很有前途的潜在治疗靶点。
Objectives Low back pain becomes a common orthopaedic disease today. It is mainly induced by the degeneration of the intervertebral disc. In this study, we tried to reveal the pathogenesis of the degeneration and the relative therapeutic strategy, which are still elusive. Materials and Methods We collected 15 degenerative intervertebral tissues and five healthy donors. Nucleus pulposus and annulus fibrosus cells were subcultured. miR-640 expression was determined by qPCR. Computer analysis and luciferase reporter assay were used to confirm miR-640 target genes. Immunohistochemical and immunocytochemical staining was used to trace the proinflammatory cytokines and key transductor of signalling pathways. We also used beta-galactosidase staining, flow cytometry, and cell viability assay to monitor the degenerative index. Results miR-640 overexpressed in patients derived degenerative nucleus pulposus tissues and cells. The inflammatory environment promoted miR-640 expression via NF-kappa B signalling pathway. In addition, miR-640 targeted to LRP1 and enhances NF-kappa B signal activity, which built a positive feedback loop. miR-640 inhibited the expression of beta-catenin and EP300, therefore, restrained WNT signal and induced the degeneration in nucleus pulposus cells. miR-640 inhibitor treatment exhibited the effects of anti-inflammation, reverse WNT signalling pathway exhaustion, and remission of degenerative characteristics in vitro. Conclusions miR-640 plays an important role in the degeneration of intervertebral disc and the relative inflammatory microenvironment. It is a promising potential therapeutic target for the low back pain biotherapy.