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Legumain promotes fibrogenesis in chronic pancreatitis via activation of transforming growth factor β1

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Abstract

Chronic pancreatitis (CP) is a major risk factor for pancreatic cancer; however, little is known about the pathogenic mechanisms underlying the development of CP. Legumain (Lgmn) has been linked to some chronic inflammatory diseases. The present study investigated the role of legumain in pancreatic fibrogenesis. We induced CP in wild type C57BL6 (WT), Lgmn-deficient (Lgmn−/−), Lgmnflox/flox and Lgmnflox/flox × LysMCre mice by intraperitoneal injection of caerulein for 4 weeks. Pancreata were collected and analyzed by quantitative reverse transcription polymerase chain reaction, Western blotting, and histology. Pancreatic stellate cells and macrophages were isolated and studied using immunofluorescence, gelatin zymography, and enzyme-linked immunosorbent assay. The effects of inhibition of legumain were investigated in vivo by administration of the specific legumain inhibitor, RR-11a. Legumain was found to be upregulated in the serum and pancreatic tissues of mice with caerulein-induced CP. Mice with global and macrophage-specific legumain deficiency exhibited significantly reduced development of pancreatic fibrosis compared with control mice, based on pancreas size, histology, and expression of fibrosis-associated genes. Our results indicate that legumain promotes activation of pancreatic stellate cells and increases synthesis of extracellular matrix proteins via activation of matrix metalloproteinase-2(MMP-2), which hydrolyzes the transforming growth factor-β1 (TGF-β1) precursor to form active TGF-β1. Administration of RR-11a markedly attenuated pancreatic fibrosis in mice with CP. Deficiency or inhibition of legumain significantly reduces the severity of pancreatic fibrosis by suppressing activation of the TGF-β1 precursor. Our results highlight the potential of legumain as a novel therapeutic target for CP.

Key messages

• Legumain expression was markedly upregulated in CP mice.

• Deletion of legumain attenuated pancreatic fibrosis in CP mice.

• Legumain promotes fibrosis via MMP-2 activation, which hydrolyzed the TGF-β1 precursor to the active form.

• Legumain is a potential therapeutic target for the management of CP.

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Acknowledgments

We thank Professor Ankang Lyu (Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China) for providing Lgmn−/−, Lgmnflox/flox, and LysMCre mice for this study. We would also like to thank Dr. Peiyuan Bai (Department of Cardiology, Zhongshan Hospital, Fudan University, Shanghai, China) for valuable suggestions and technical assistance on our experiments.

Funding

This work was sponsored by Natural Science Foundation of China grants to Gongyong Hu (81670584 and 81970556) and Xingpeng Wang (81570580), and the Shanghai Pujiang Program to Gongyong Hu (18PJD041).

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Author notes

  1. Ying-Chun Ren, Qiuyan Zhao and Yan He contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai, 200080, China

    Ying-Chun Ren, Qiuyan Zhao, Yan He, Bin Li, Zengkai Wu, Juanjuan Dai, Li Wen, Xingpeng Wang & Guoyong Hu

  2. Shanghai Key Laboratory of Pancreatic Disease, Institute of Pancreatic Disease, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China

    Ying-Chun Ren, Qiuyan Zhao, Yan He, Bin Li, Zengkai Wu, Juanjuan Dai, Li Wen, Xingpeng Wang & Guoyong Hu

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  1. Ying-Chun Ren
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Correspondence to Xingpeng Wang or Guoyong Hu.

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All animal experiments were performed in accordance with the principle for replacement, refinement, and reduction (the 3Rs) and were approved by the Animal Ethics Committee of Shanghai Jiao Tong University School of Medicine (SYXK 2013-0050, Shanghai, China.).

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Ren, YC., Zhao, Q., He, Y. et al. Legumain promotes fibrogenesis in chronic pancreatitis via activation of transforming growth factor β1. J Mol Med 98, 863–874 (2020). https://doi.org/10.1007/s00109-020-01911-0

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