Renal Unit, Department of Medicine, University-Hospital of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
Department of General Surgery and Odontoiatrics, Liver Transplant Unit, University Hospital of Verona, Piazzale Stefani 1, 37126, Verona, Italy.
J Transl Med. 2019 Jan 7;17(1):12. doi: 10.1186/s12967-019-1770-1.
The epithelial to mesenchymal transition (EMT) is a multi-factorial biological mechanism involved in renal and hepatic fibrosis and the IL-1 beta has been assumed as a mediator of this process although data are not exhaustive. Therefore, the aim of our study was to evaluate the role of this cytokine in the EMT of renal proximal tubular epithelial cells (HK-2) and stellate cells (LX-2) and the protective/anti-fibrotic effect of its inhibition by Canakinumab (a specific human monoclonal antibody targeted against IL-1beta).
Both cell types were treated with IL-1 beta (10 ng/ml) for 6 and 24 h with and without Canakinumab (5 μg/ml). As control we used TGF-beta (10 ng/ml). Expression of EMT markers (vimentin, alpha-SMA, fibronectin) were evaluated through western blotting and immunofluorescence. Genes expression for matrix metalloproteinases (MMP)-2 was measured by Real-Time PCR and enzymatic activity by zymography. Cellular motility was assessed by scratch assay.
IL-1 beta induced a significant up-regulation of EMT markers in both cell types and increased the MMP-2 protein expression and enzymatic activity, similarly to TGF-beta. Moreover, IL-1 beta induced a higher rate of motility in HK-2. Canakinumab prevented all these modifications in both cell types.
Our results clearly demonstrate the role of IL-1 beta in the EMT of renal/stellate cells and it underlines, for the first time, the therapeutic potential of its specific inhibition on the prevention/minimization of organ fibrosis.
上皮间质转化（EMT）是一个多因素的生物学机制，涉及到肾脏和肝脏纤维化，IL-1β 被认为是这个过程的介质，尽管数据并不详尽。因此，我们的研究目的是评估这种细胞因子在肾近端肾小管上皮细胞（HK-2）和星状细胞（LX-2）的 EMT 中的作用，以及其抑制物 Canakinumab（一种针对 IL-1β 的特异性人单克隆抗体）的保护/抗纤维化作用。
用 IL-1β（10ng/ml）处理两种细胞类型 6 和 24 小时，同时用和不用 Canakinumab（5μg/ml）。我们使用 TGF-β（10ng/ml）作为对照。通过 Western blot 和免疫荧光法评估 EMT 标志物（波形蛋白、α-SMA、纤维连接蛋白）的表达。通过实时 PCR 测量基质金属蛋白酶（MMP）-2 的基因表达，通过酶谱法测量酶活性。通过划痕实验评估细胞迁移率。
IL-1β 在两种细胞类型中都显著上调了 EMT 标志物的表达，并增加了 MMP-2 蛋白表达和酶活性，与 TGF-β 相似。此外，IL-1β 还增加了 HK-2 的迁移率。Canakinumab 预防了这两种细胞类型的所有这些变化。
我们的结果清楚地表明了 IL-1β 在肾/星状细胞 EMT 中的作用，并首次强调了其特异性抑制在预防/最小化器官纤维化中的治疗潜力。
