Pro-inflammatory Cytokines Drive Deregulation of Potassium Channel Expression in Primary Synovial Fibroblasts
Pro-inflammatory Cytokines Drive Deregulation of Potassium Channel Expression in Primary Synovial Fibroblasts
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DOI：
10.3389/fphys.2020.00226
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发表时间：
2020-03-24
影响因子：
4
通讯作者：
Barrett-Jolley, Richard
中科院分区：
文献类型：
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作者：
Haidar, Omar;O'Neill, Nathanael;Barrett-Jolley, Richard
滑膜分泌滑液，同时也富含伤害感受器，并且是无血管关节组织与循环系统之间的通道。在关节炎模型中，驻留的成纤维细胞样滑膜细胞（FLS）的钙激活钾通道（K-Ca）活性会发生变化，且这种变化与FLS的激活相关。目的：在炎症性关节炎体外模型中研究这种激活情况，采用细胞因子肿瘤坏死因子α（TNF-α）和白细胞介素1β（IL-1β）处理72小时。方法：从大鼠滑膜中分离FLS细胞。我们通过RNA测序分析FLS mRNA的整体变化，随后聚焦于FLS离子通道基因以及相应的FLS电生理表型，最后利用Ingenuity通路分析（IPA）和MATLAB对数据进行建模。结果：IPA显示细胞因子显著激活了炎症、骨关节炎和钙信号经典通路，我们鉴定出约200种通道基因转录本。大电导钙激活钾通道（BK）由形成孔道的Kcnma1以及β亚基组成。细胞因子处理后，通过定量聚合酶链反应（qPCR）检测到Kcnma1 RNA丰度显著增加，通过RNA测序检测到多个离子通道发生变化，包括BK通道β亚基Kcnmb1/2表达缺失以及Kcnmb3表达增加。在电生理实验中，20毫伏时总体电流密度降低，但该电位下弦电导无变化。结论：在体外对FLS进行TNF-α和IL-1β处理，在转录组水平上重现了炎症性关节炎的几个常见特征，包括Kcnma1和Kcnmb3基因表达增加。
The synovium secretes synovial fluid, but is also richly innervated with nociceptors and acts as a gateway between avascular joint tissues and the circulatory system. Resident fibroblast-like synoviocytes' (FLS) calcium-activated potassium channels (K-Ca) change in activity in arthritis models and this correlates with FLS activation. Objective To investigate this activation in an in vitro model of inflammatory arthritis; 72 h treatment with cytokines TNF alpha and IL1 beta. Methods FLS cells were isolated from rat synovial membranes. We analyzed global changes in FLS mRNA by RNA-sequencing, then focused on FLS ion channel genes and the corresponding FLS electrophysiological phenotype and finally modeling data with ingenuity pathway analysis (IPA) and MATLAB. Results IPA showed significant activation of inflammatory, osteoarthritic and calcium signaling canonical pathways by cytokines, and we identified similar to 200 channel gene transcripts. The large K-Ca (BK) channel consists of the pore forming Kcnma1 together with beta-subunits. Following cytokine treatment, a significant increase in Kcnma1 RNA abundance was detected by qPCR and changes in several ion channels were detected by RNA-sequencing, including a loss of BK channel beta-subunit expression Kcnmb1/2 and an increase in Kcnmb3. In electrophysiological experiments, there was a decrease in over-all current density at 20 mV without change in chord conductance at this potential. Conclusion TNF alpha and IL1 beta treatment of FLS in vitro recapitulated several common features of inflammatory arthritis at the transcriptomic level, including increase in Kcnma1 and Kcnmb3 gene expression.