De Paepe Boel, Martin Jean-Jacques, Herbelet Sandrine, Jimenez-Mallebrera Cecilia, Iglesias Estibaliz, Jou Cristina, Weis Joachim, De Bleecker Jan L
Department of Neurology, Neuromuscular Reference Center, Ghent University Hospital, Ghent, Belgium.
Department of Ultrastructural Neuropathology, Born-Bunge Institute, Antwerp University Hospital, Wilrijk, Belgium.
Lab Invest. 2016 Aug;96(8):872-84. doi: 10.1038/labinvest.2016.68. Epub 2016 Jun 20.
Alongside well-known nuclear factor κB (NFκB) and its associated cytokine networks, nuclear factor of activated T cells 5 (NFAT5), the master regulator of cellular osmoprotective programs, comes forward as an inflammatory regulator. To gain insight into its yet unexplored role in muscle disease, we studied the expression of NFAT5 target proteins involved in osmolyte accumulation: aldose reductase (AR), taurine transporter (TauT), and sodium myo-inositol co-transporter (SMIT). We analyzed idiopathic inflammatory myopathy and Duchenne muscular dystrophy muscle biopsies and myotubes in culture, using immunohistochemistry, immunofluorescence, and western blotting. We report that the level of constitutive AR was upregulated in patients, most strongly so in Duchenne muscular dystrophy. TauT and SMIT expression levels were induced in patients' muscle fibers, mostly representing regenerating and atrophic fibers. In dermatomyositis, strong staining for AR, TauT, and SMIT in atrophic perifascicular fibers was accompanied by staining for other molecular NFAT5 targets, including chaperones, chemokines, and inducible nitric oxide synthase. In these fibers, NFAT5 and NFκB p65 staining coincided, linking both transcription factors with this important pathogenic hallmark. In sporadic inclusion body myositis, SMIT localized to inclusions inside muscle fibers. In addition, SMIT was expressed by a substantial subset of muscle-infiltrating macrophages and T cells in patient biopsies. Our results indicate that osmolyte pathways may contribute to normal muscle functioning, and that activation of AR, TauT, and SMIT in muscle inflammation possibly contributes to the tissue's failing program of damage control.
除了著名的核因子κB(NFκB)及其相关的细胞因子网络外,活化T细胞核因子5(NFAT5)作为细胞渗透保护程序的主要调节因子,也成为一种炎症调节因子。为了深入了解其在肌肉疾病中尚未被探索的作用,我们研究了参与渗透溶质积累的NFAT5靶蛋白的表达:醛糖还原酶(AR)、牛磺酸转运体(TauT)和钠-肌醇共转运体(SMIT)。我们使用免疫组织化学、免疫荧光和蛋白质印迹法分析了特发性炎性肌病和杜兴氏肌营养不良症的肌肉活检样本以及培养的肌管。我们报告称,患者体内组成型AR的水平上调,在杜兴氏肌营养不良症患者中上调最为明显。TauT和SMIT的表达水平在患者的肌纤维中被诱导,主要代表再生和萎缩的肌纤维。在皮肌炎中,萎缩的束周纤维中AR、TauT和SMIT的强染色伴随着其他分子NFAT5靶标的染色,包括伴侣蛋白、趋化因子和诱导型一氧化氮合酶。在这些纤维中,NFAT5和NFκB p65染色重合,将这两种转录因子与这一重要的致病特征联系起来。在散发性包涵体肌炎中,SMIT定位于肌纤维内的包涵体。此外,在患者活检样本中,相当一部分浸润肌肉的巨噬细胞和T细胞表达SMIT。我们的结果表明,渗透溶质途径可能有助于正常肌肉功能,并且肌肉炎症中AR、TauT和SMIT的激活可能导致组织损伤控制程序的失败。