Abstract：
The prevalence of chronic kidney disease (CKD) in dogs increases with age, and renal fibrosis is an important pathophysiological mechanism in this process. However, only a few drugs that can effectively inhibit fibrosis in the kidneys of dogs are currently available. In this study, we aimed to determine whether pirfenidone, a drug that has shown antifibrotic effects in various clinical studies, also exerts antifibrotic effects on canine renal tubular epithelial cells, Madin-Darby canine kidney cells (MDCK). To this end, we treated MDCK cells with various concentrations of pirfenidone, followed by transforming growth factor-beta1 (TGF-β1) to stimulate fibrotic conditions. A cell viability assay was performed to determine the effect of pirfenidone on cell survival. Fibrosis-related markers and TGF-β1 fibrotic pathway-related markers were assessed using qPCR, Western blot analysis and immunocytochemistry. A one-way analysis of variance (ANOVA) was performed, followed by Tukey\'s post-hoc test for multiple comparisons. Pirfenidone treatment significantly reduced the expression of profibrotic markers such as α-smooth muscle actin, fibronectin, and collagen. Additionally, it upregulated the expression of E-cadherin, an epithelial marker. Furthermore, pirfenidone effectively inhibited the phosphorylation of key factors involved in the TGF-β1 signaling pathway, including Smad2/3 and ERK1/2. These results demonstrate that pirfenidone suppresses TGF-β1-induced fibrosis in MDCK cells by attenuating epithelial-mesenchymal transition and the relevant signaling pathways.
摘要：
犬慢性肾脏病(CKD)患病率随年龄增长而增加,肾纤维化是这一过程中重要的病理生理机制。然而,目前只有少数能有效抑制犬肾纤维化的药物。在这项研究中,我们的目的是确定吡非尼酮，一种在各种临床研究中显示出抗纤维化作用的药物，还对犬肾小管上皮细胞发挥抗纤维化作用，Madin-Darby犬肾细胞(MDCK)。为此,我们用各种浓度的吡非尼酮处理MDCK细胞，然后用转化生长因子-β1（TGF-β1）刺激纤维化条件。进行细胞活力测定以确定吡非尼酮对细胞存活的影响。纤维化相关标志物和TGF-β1纤维化途径相关标志物使用qPCR评估，蛋白质印迹分析和免疫细胞化学。进行了单向方差分析(ANOVA)，其次是Tukey的事后多重比较测试。吡非尼酮治疗显着降低促纤维化标志物如α-平滑肌肌动蛋白的表达，纤连蛋白,和胶原蛋白。此外,它上调了E-cadherin的表达，上皮标记物.此外，吡非尼酮能有效抑制TGF-β1信号通路中关键因子的磷酸化,包括Smad2/3和ERK1/2。这些结果表明，吡非尼酮通过减弱上皮-间质转化和相关信号通路来抑制TGF-β1诱导的MDCK细胞纤维化。
