IL-6-accelerated calcification by induction of ROR2 in human adipose tissue-derived mesenchymal stem cells is STAT3 dependent
IL-6-accelerated calcification by induction of ROR2 in human adipose tissue-derived mesenchymal stem cells is STAT3 dependent
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DOI：
10.1093/rheumatology/ket496
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发表时间：
2014-07-01
期刊：
影响因子：
5.5
通讯作者：
Tanaka, Yoshiya
中科院分区：
文献类型：
--
作者：
Fukuyo, Shunsuke;Yamaoka, Kunihiro;Tanaka, Yoshiya
方法.使用在成骨细胞诱导培养基中培养的hADSC评估炎性细胞因子的作用。通过实时PCR测量mRNA表达，并通过蛋白质印迹法测量蛋白质水平。茜素红S染色检测细胞矿化情况。在hADSC中，IL-6/可溶性IL-6受体（sIL-6 R）、TNF或IL-1 β通过增强成骨细胞分化标志物Runt相关转录因子2（RUNX 2）的表达加速钙化。IL-6/sIL-6 R的作用最大。受体酪氨酸激酶样孤儿受体2（ROR 2），参与非经典无翅型（WNT）MMTV整合位点途径的mRNA转录增加，而β-连环蛋白表达，成骨细胞分化的经典WNT信号通路中的一个重要因素，没有改变。小干扰RNA（siRNA）抑制信号转导子和转录激活子3（STAT 3）而非STAT 1，对RUNX 2和ROR 2的表达产生强烈的抑制作用，并抑制加速钙化。IL-6/sIL-6 R刺激以STAT 3依赖性方式加速hADSC中的ROR 2/WNT 5A通路，导致钙化增强。这些结果表明，hADSCs中IL-6加速异位钙化的机制可能涉及慢性炎症组织，IL-6抑制剂可能有助于治疗炎症性疾病中的异位钙化。
Methods. The effects of inflammatory cytokines were evaluated using hADSCs cultured in osteoblast induction medium. mRNA expression was measured by real-time PCR and protein levels were measured by western blotting. Cell mineralization was evaluated by Alizarin Red S staining.Results. In hADSCs, administration of IL-6/soluble IL-6 receptor (sIL-6R), TNF or IL-1 beta accelerated calcification through enhanced expression of an osteoblast differentiation marker, runt-related transcription factor 2 (RUNX2). IL-6/sIL-6R had the greatest effect. The transcription of mRNA for receptor tyrosine kinase-like orphan receptor 2 (ROR2), involved in the non-canonical wingless-type (WNT) MMTV integration site pathway, was increased, while beta-catenin expression, an essential factor in the canonical WNT signalling pathway for osteoblast differentiation, did not change. Suppression of signal transducer and activator of transcription 3 (STAT3), but not STAT1, by small interfering RNA (siRNA) exerted a strong inhibitory effect on RUNX2 and ROR2 expression, and inhibited accelerated calcification.Conclusion. IL-6/sIL-6R stimulation accelerated the ROR2/WNT5A pathway in hADSCs in a STAT3-dependent manner, resulting in augmented calcification. These results suggest that the mechanisms of ectopic calcification accelerated by IL-6 in hADSCs may be involved in chronic inflammatory tissues and that IL-6 inhibitors may be beneficial in the treatment of ectopic calcification in inflammatory diseases.