O'Reilly Steven, Ciechomska Marzena, Fullard Nicola, Przyborski Stefan, van Laar Jacob M
Faculty of Health and Life Sciences, Northumbria University, Newcastle Upon Tyne, NE1 8ST, United Kingdom.
Institute of Cellular Medicine Faculty of Medical Sciences, MRG 4th Floor Cookson Building Framlington Place, Newcastle Upon Tyne, NE2 4HH.
Sci Rep. 2016 Apr 26;6:25066. doi: 10.1038/srep25066.
Systemic sclerosis is an autoimmune connective tissue disease in which T cells play a prominent role. We and others have previously demonstrated a role for T cell-derived IL-13 in mediating the induction of collagen in dermal fibroblasts and that blockade with IL-13 antibodies attenuates this increase. In this study we want to probe the signalling that underpins IL-13 mediated matrix deposition. Isolated dermal fibroblasts were incubated with recombinant IL-13 and gene expression by qRT-PCR was performed for collagen1A1 and TGF-β1. Small interfering RNA (siRNA) was used to knock down STAT6 and a small molecule inhibitor was also used to block this pathway. MiR-135b was transfected into fibroblasts plus and minus IL-13 to see if this miR plays a role. miR-135b was measured in systemic sclerosis fibroblasts isolated from patients and also in serum. Results showed that IL-13 increased collagen expression and that this is independent from TGF-β1. This is dependent on STAT6 as targeting this blocked induction. MiR-135b reduces collagen induction in fibroblasts and scleroderma fibroblasts have lower constitutive levels of the miR. We further demonstrate that miR135b is repressed by methylation and may include MeCP2. In conclusion we show that STAT6 and miR-135b regulate IL-13-mediated collagen production by fibroblasts.
系统性硬化症是一种自身免疫性结缔组织疾病，其中T细胞发挥着重要作用。我们和其他人之前已经证明，T细胞衍生的白细胞介素-13（IL-13）在介导真皮成纤维细胞中胶原蛋白的诱导过程中发挥作用，并且用IL-13抗体进行阻断可减弱这种增加。在本研究中，我们想要探究支撑IL-13介导的基质沉积的信号传导。将分离的真皮成纤维细胞与重组IL-13一起孵育，并通过qRT-PCR对胶原蛋白1A1和转化生长因子-β1（TGF-β1）进行基因表达检测。使用小分子干扰RNA（siRNA）敲低信号转导和转录激活因子6（STAT6），并且还使用小分子抑制剂来阻断该途径。将miR-135b转染到添加和不添加IL-13的成纤维细胞中，以观察该miR是否发挥作用。在从患者分离的系统性硬化症成纤维细胞以及血清中测量miR-135b。结果表明，IL-13增加了胶原蛋白的表达，并且这与TGF-β1无关。这依赖于STAT6，因为靶向它会阻断诱导。miR-135b降低了成纤维细胞中胶原蛋白的诱导，并且硬皮病成纤维细胞中该miR的组成水平较低。我们进一步证明，miR135b被甲基化抑制，并且可能包括甲基化CpG结合蛋白2（MeCP2）。总之，我们表明STAT6和miR-135b调节成纤维细胞中IL-13介导的胶原蛋白产生。
