To explore the role of Notch1 pathway in the pathogenesis of podocyte injury, and to provide novel strategy for podocyte repair in lupus nephritis (LN). Bioinformatics analysis and immunofluorescence assay were applied to determine the expression and localization of Notch1 intracellular domain1 (NICD1) in kidneys of LN patients and MRL/lpr mice. The stable podocyte injury model in vitro was established by puromycin aminonucleoside (PAN) treatment. Expression of inflammasome activation related gene was detected by qPCR. The podocytes with PAN treatment were cultured with or without N-S-phenyl-glycine-t-butylester (DAPT), an inhibitor of Notch1 pathway. NICD1, Wilm’stumor1 (WT1), nucleotide-binding oligomerization domain-like receptors 3 (NLRP3), and absent in melanoma-like receptors 2 (AIM2) were detected by western blot. In vivo, MRL/lpr mice were administrated with DAPT or vehicle. The LN symptoms were assessed. The podocyte injury was evaluated, and the NLRP3 in podocytes of mice was detected. Notch1 pathway was overactivated in glomeruli of LN patients. NICD1 was colocalized with podocytes of LN patients and MRL/lpr mice. The inflammasome-related genes were significantly increased in podocytes with PAN treatment. NICD1 and NLRP3 were significantly decreased, while WT1 was significantly increased in injured podocytes treated with DAPT in vitro. In vivo, lupus-like symptoms were alleviated in DAPT treatment group. Notch1 pathway was inhibited in kidneys of mice treated with DAPT. The renal inflammation was reduced and the podocyte injury was mitigated in DAPT treatment group. The NLRP3 was decreased in podocytes of mice treated with DAPT. Notch1 pathway was overactivated in podocytes of LN patients and MRL/lpr mice. Blockade of Notch1 pathway reduced renal inflammation and alleviated podocyte injury via inhibition of NLRP3 inflammasome activation in LN.

探讨Notch1通路在狼疮性肾炎（LN）足细胞损伤发病机制中的作用，为狼疮性肾炎（LN）足细胞修复提供新策略。应用生物信息学分析和免疫荧光法测定Notch1胞内域1（NICD1）在LN患者和MRL/lpr小鼠肾脏中的表达和定位。采用嘌呤霉素氨基核苷（PAN）处理建立稳定的体外足细胞损伤模型。qPCR检测炎症小体激活相关基因的表达。用 PAN 处理的足细胞在有或没有 NS-苯基甘氨酸叔丁酯 (DAPT)（一种 Notch1 通路抑制剂）的情况下培养。通过蛋白质印迹检测到 NICD1、Wilm'stumor1 (WT1)、核苷酸结合寡聚化结构域样受体 3 (NLRP3) 和黑色素瘤样受体 2 (AIM2) 中缺失的蛋白。在体内，给MRL/lpr小鼠施用DAPT或媒介物。评估 LN 症状。评估足细胞损伤情况，并检测小鼠足细胞中的NLRP3。LN 患者肾小球中 Notch1 通路过度激活。NICD1 与 LN 患者和 MRL/lpr 小鼠的足细胞共定位。PAN 处理后足细胞中炎症小体相关基因显着增加。体外经 DAPT 处理的损伤足细胞中 NICD1 和 NLRP3 显着降低，而 WT1 显着增加。在体内，DAPT治疗组狼疮样症状得到缓解。DAPT 治疗小鼠肾脏中的 Notch1 通路受到抑制。DAPT治疗组肾脏炎症减轻，足细胞损伤减轻。经 DAPT 处理的小鼠足细胞中 NLRP3 减少。LN 患者和 MRL/lpr 小鼠的足细胞中 Notch1 通路过度激活。阻断 Notch1 通路可通过抑制 LN 中 NLRP3 炎性体激活来减少肾脏炎症并减轻足细胞损伤。