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Blockade of Notch1 Signaling Alleviated Podocyte Injury in Lupus Nephritis Via Inhibition of NLRP3 Inflammasome Activation

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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.

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Acknowledgements

We thank Dr. Jiaheng Xie, for his kind suggestion in the revision of the manuscript.

Funding

This work was supported by the National Key R&D Program of China (2020YFA0710800), the Key Program of National Natural Science Foundation of China (81930043), the Jiangsu Provincial Key Research and Development Program (BE2020621) and the National Science Foundation of Jiangsu Province (BK20210016).

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Authors and Affiliations

  1. Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210008, China

    Dan Wu, Shiyi Zhang, Ying Zhu, Dongdong Zhang, Honghong Yu, Genhong Yao & Lingyun Sun

  2. Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing, 210008, China

    Tingting Jiang, Mengxi Huang, Yuanyuan Zheng & Lingyun Sun

  3. Department of Hepatobiliary and Pancreatic Surgery, Conversion therapy center for Hepatobiliary and Pancreatic Tumors, First Hospital of Jiaxing, Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang, People’s Republic of China

    Liang Chen

  4. Department of Rheumatology and Immunology, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, People’s Republic of China

    Lingyun Sun

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  1. Dan Wu
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Contributions

All authors contributed to the study conception and design. WD, YGH and SLY designed experiments. WD and JTT carried out experiments. Data collection and analysis were performed by WD, ZSY, HMX, ZY and CL. The first draft of the manuscript was written by WD. All authors commented on previous versions of the manuscript. ZYY, ZDD and YHH performed the language modifying. All authors read and approved the final manuscript.

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Correspondence to Genhong Yao or Lingyun Sun.

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This study was performed in line with the principles of the Declaration of Helsinki, and approved by the Ethics Committee of Nanjing Drum Tower Hospital. This study followed the NIH Guide for the Care and Use of Laboratory Animals. All animal experimental protocols were approved by the Ethics Committee of Nanjing Drum Tower Hospital.

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Wu, D., Jiang, T., Zhang, S. et al. Blockade of Notch1 Signaling Alleviated Podocyte Injury in Lupus Nephritis Via Inhibition of NLRP3 Inflammasome Activation. Inflammation 47, 649–663 (2024). https://doi.org/10.1007/s10753-023-01935-x

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