Inhibition of cellular communication network factor 1 (CCN1)-driven senescence slows down cartilage inflammaging and osteoarthritis
Inhibition of cellular communication network factor 1 (CCN1)-driven senescence slows down cartilage inflammaging and osteoarthritis
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DOI：
10.1016/j.bone.2020.115522
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发表时间：
2020-10-01
期刊：
影响因子：
4.1
通讯作者：
Qiu, Yusheng
中科院分区：
文献类型：
--
作者：
Feng, Meng;Peng, Hang;Qiu, Yusheng
目的：为探讨细胞通讯网络因子1（cellularcommunicationnetworkfactor 1，CCN 1）在软骨炎症和骨关节炎（osteoarthritis，OA）发病机制中的作用，在体外分离的原代人软骨细胞、离体软骨组织块和临床前小鼠模型中进行研究。提取RNA以定量分解代谢靶标和促炎基因，并用特异性抗体探测蛋白质。通过ELISA监测IL-1 β和IL-6。进行IHC以评价重要的肥大标志物和分解代谢标志物。丹参酮Ⅱ A对软骨细胞的作用呈时间依赖性和剂量依赖性。将锦鸡儿外植体在生长培养基中培养，并进一步用丹参酮IIA处理。在13月龄C57 BL/6 J小鼠中进行关节内注射。采用番红O染色和快速绿色染色观察软骨组织学变化，并采用OARSI评分系统进行半定量分析。骨关节炎病变区CCN 1表达较非病变区明显增加，与骨关节炎的病理改变密切相关。CCN 1的过表达促进软骨细胞衰老，而小干扰RNA下调CCN 1的表达可减少CCN 1的产生并限制炎症分泌，提示CCN 1可能是干预OA的新靶点。丹参酮IIA抑制CCN 1可减少SASP组分，且呈剂量和时间依赖性。此外，我们的数据表明，丹参酮IIA能够保持关节软骨完整性，抑制CCN 1的产生，并抑制SASP因子在人软骨外植体和老年小鼠模型中。结论：本研究表明，CCN 1信号通路加重软骨炎症和基质降解。总的来说，我们的研究结果显示了通过抑制CCN 1轴来重新利用丹参酮IIA减缓人类和小鼠OA进展的新见解。
Objective: To explore the role of cellular communication network factor 1 (CCN1) in cartilage inflammaging and osteoarthritis (OA) pathogenesis in the isolated primary human chondrocytes in vitro, cartilage explants ex vivo, and a pre-clinical mice model.Methods: Recombinant human CCN1 stimulation and small interfering RNA inhibition were conducted in human chondrocytes. The RNA was extracted to quantify catabolic targets and pro-inflammatory genes and the proteins were probed with specific antibodies. IL-1 ss and IL-6 were monitored by ELISA. IHC was performed to evaluate important hypertrophic hallmarks and catabolic markers. The effects of Tanshinone IIA on chondrocytes were investigated in both time-dependent and dose-dependent processes. Cartilage explants were cultured in growth medium and further treated with Tanshinone IIA. The intra-articular injection was performed in 13 months old C57BL/6J mice. Safranin O and fast green staining were performed to evaluate the histological change of cartilage followed by a semi-quantitative analysis using the OARSI scoring system.Results: RNA and protein levels of CCN1 increased in an age-dependent manner compared to young donors. Increased CCN1 expression was also found in the damaged area compared to the non-lesion area which correlated with the advanced pathological change in human OA. The overexpression of CCN1 promoted chondrocytes senescence, while the down-regulation of CCN1 by small interfering RNA reduced CCN1 production and limited inflammation secretion suggesting that CCN1 was a possible novel target to intervene OA. Inhibition of CCN1 by using Tanshinone IIA could reduce SASP components in a dose- and time-dependent manner. Additionally, our data showed that Tanshinone IIA was able to preserve articular cartilage integrity, suppress CCN1 production, and inhibit SASP factors in human cartilage explants and in aged mice model.Conclusion: This study showed that CCN1 signaling aggravated cartilage inflammaing and matrix degradation. Collectively, our findings showed new insight into repurposing Tanshinone IIA for slowing down OA advancement in human and mice by inhibiting the CCN1 axis.