Center of Kidney Dialysis, The Shaanxi Provincial People's Hospital, Xi'an, 710068, China; Shaanxi University of Chinese Medicine, Xi'an, 712046, China.
Shaanxi University of Chinese Medicine, Xi'an, 712046, China.
Biochem Biophys Res Commun. 2019 Dec 3;520(2):413-419. doi: 10.1016/j.bbrc.2019.09.130. Epub 2019 Oct 11.
Peritoneal fibrosis (PF) caused by long-term peritoneal dialysis is closely associated with the epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs). Moreover, the anti-fibrotic role of Arctigenin (Arc) has been reported in several fibrosis disorders. Therefore, the preventive effect of Arc on transforming growth factor-β1 (TGF-β1)-induced EMT and the underlying mechanisms in HPMCs was investigated in this study. Firstly, the PD model was established by TGF-β1 stimulation in cultured HPMCs in vitro, we found that TGF-β1 significantly increased the EMT markers (α-SMA, vimentin, and fibronectin) and plasminogen activator inhibitor type 1 (PAI-1) expressions, but decreased epithelial marker (E-cadherin). Co-treatment with Arc (10, 20, 40 μM) ameliorated TGF-β1-induced EMT in a dose-dependent manner, and the expression of PAI-1 was also inhibited by Arc, which was abrogated by restoration of PAI-1. Moreover, Arc enhanced the phosphorylated AMP-activated protein kinase (AMPK), but inhibited the phosphorylated IκBα level and nuclear translocation of nuclear factor κB (NF-κB) p65 in TGF-β1-induced HPMCs. ChIP and Luciferase reporter assays verified that the increased binding capacity of NF-κB to the promoter of PAI-1 induced by TGF-β1 was reversely attenuated by Arc in HPMCs. However, the effect of Arc on TGF-β1-induced NF-κB activation, PAI-1 expression and EMT in HPMCs was attenuated by AMPK agonist Compound C. In conclusion, these data demonstrated that Arc suppressed TGF-β1-induced EMT by activating the AMPK/NF-κB pathway to inhibit PAI-1 expression in HPMCs. Therefore, Arc might act as a potential therapeutic agent for PD treatment.
腹膜纤维化(PF)是由长期腹膜透析引起的,与人类腹膜间皮细胞(HPMCs)的上皮-间充质转化(EMT)密切相关。此外,已报道牛蒡子苷(Arc)在几种纤维化疾病中具有抗纤维化作用。因此,本研究旨在探讨 Arc 对 TGF-β1 诱导的 EMT 及 EMT 相关机制在 HPMCs 中的预防作用。首先,我们通过 TGF-β1 刺激体外培养的 HPMCs 建立 PD 模型,发现 TGF-β1 可显著增加 EMT 标志物(α-SMA、波形蛋白和纤连蛋白)和纤溶酶原激活物抑制剂 1(PAI-1)的表达,但降低上皮标志物(E-钙粘蛋白)的表达。Arc(10、20、40 μM)以剂量依赖性方式改善 TGF-β1 诱导的 EMT,Arc 还抑制了 PAI-1 的表达,而 PAI-1 的恢复则消除了这种抑制作用。此外,Arc 增强了磷酸化 AMP 激活蛋白激酶(AMPK),但抑制了 TGF-β1 诱导的 HPMCs 中磷酸化 IκBα水平和核转录因子κB(NF-κB)p65的核转位。ChIP 和荧光素酶报告基因分析验证了 Arc 在 HPMCs 中可逆转 TGF-β1 诱导的 PAI-1 启动子与 NF-κB 结合能力的增加。然而,Arc 对 TGF-β1 诱导的 NF-κB 激活、PAI-1 表达和 EMT 的作用在 HPMCs 中被 AMPK 激动剂 Compound C 减弱。综上所述,这些数据表明,Arc 通过激活 AMPK/NF-κB 通路抑制 HPMCs 中 PAI-1 的表达,从而抑制 TGF-β1 诱导的 EMT。因此,Arc 可能是治疗 PD 的一种潜在治疗剂。