Department of Nephrology, Tongren Hospital of Wuhan University (Wuhan Third Hospital), Wuhan University, Wuhan, Hubei 430074, PR China.
Department of Nephrology, Huadu District People's Hospital, Southern Medical University, Guangzhou, Guangdong 510800, PR China.
Int Immunopharmacol. 2019 Jan;66:198-204. doi: 10.1016/j.intimp.2018.11.027. Epub 2018 Nov 22.
It has been demonstrated that thioredoxin-interacting protein (TXNIP) interacted with NACHT, LRR and PYD domains-containing protein 3 (NLRP3) and participated in the NLRP3 inflammasome activation. Our previous study has demonstrated that in human peritoneal mesothelial cells (HPMCs), exposure to high glucose-based peritoneal dialysis (PD) solutions induced mitochondrial reactive oxygen species (ROS) production, activation of NLRP3 inflammasome and IL-1β expression. This study aimed to investigate the effect of high glucose-based PD fluids on the TXNIP expression and the underlying mechanisms by which TXNIP-NLRP3 interaction mediates the inflammatory injury to HPMCs in high glucose-based PD fluids conditions.
TXNIP gene and protein expression was detected by real-time polymerase chain reaction (RT-PCR) and immunoblot. Immunoprecipitation was used to evaluate the interaction between TRX1 and TXNIP, TXNIP and NLRP3. ROS production and IL-1β expression was examined by flow cytometry and immunoblot and enzyme-linked immunosorbent assay (ELISA) respectively.
It was identified that high glucose-based PD solutions enhance the level of TXNIP gene and protein in cultured HPMCs and a rat-based PD model. We also found that ROS generation induced by high glucose-based PD solutions disrupts the TRX1-TXNIP association, while promoting the binding of TXNIP to NLRP3 in HPMCs. Furthermore, the application of a ROS inhibitor (APDC) to HPMCs blocked the high glucose-based PD solution-induced TXNIP-NLRP3 binding, in addition to ROS production and IL-1β expression.
The results of the present study revealed a novel mechanism underlying high glucose-containing PD-mediated peritoneal inflammatory injury, supporting the attenuation of ROS generation as a potential therapeutic strategy to alleviate such pathology.
已有研究表明,硫氧还蛋白相互作用蛋白(TXNIP)与 NACHT、LRR 和 PYD 结构域包含蛋白 3(NLRP3)相互作用,并参与 NLRP3 炎性小体的激活。我们之前的研究表明,在人腹膜间皮细胞(HPMCs)中,高糖基腹膜透析(PD)溶液暴露会导致线粒体活性氧(ROS)产生、NLRP3 炎性小体的激活和 IL-1β 的表达。本研究旨在探讨高糖基 PD 液对 TXNIP 表达的影响,以及 TXNIP-NLRP3 相互作用介导高糖基 PD 液条件下 HPMCs 炎症损伤的潜在机制。
实时聚合酶链反应(RT-PCR)和免疫印迹法检测 TXNIP 基因和蛋白表达。免疫沉淀法用于评估 TRX1 与 TXNIP、TXNIP 与 NLRP3 之间的相互作用。通过流式细胞术和免疫印迹法以及酶联免疫吸附试验(ELISA)分别检测 ROS 产生和 IL-1β 表达。
研究发现,高糖基 PD 溶液增强了培养的 HPMCs 和大鼠 PD 模型中 TXNIP 基因和蛋白的水平。我们还发现,高糖基 PD 溶液诱导的 ROS 生成破坏了 TRX1-TXNIP 结合,同时促进了 TXNIP 与 HPMCs 中 NLRP3 的结合。此外,ROS 抑制剂(APDC)在 HPMCs 中的应用阻断了高糖基 PD 溶液诱导的 TXNIP-NLRP3 结合,同时抑制了 ROS 生成和 IL-1β 的表达。
本研究揭示了高糖基 PD 介导的腹膜炎症损伤的新机制,支持抑制 ROS 生成作为缓解这种病理的潜在治疗策略。