Nintedanib attenuates peritoneal fibrosis by inhibiting mesothelial‐to‐mesenchymal transition, inflammation and angiogenesis

Abstract

Nintedanib, an Food and Drug Administration (FDA) approved multiple tyrosine kinase inhibitor, exhibits an anti‐fibrotic effect in lung and kidneys. Its effect on peritoneal fibrosis remains unexplored. In this study, we found that nintedanib administration lessened chlorhexidine gluconate (CG)‐induced peritoneal fibrosis and reduced collagen I and fibronectin expression. This coincided with suppressed phosphorylation of platelet‐derived growth factor receptor, fibroblast growth factor receptors, vascular endothelial growth factor receptor and Src family kinase. Mechanistically, nintedanib inhibited injury‐induced mesothelial‐to‐mesenchymal transition (MMT), as demonstrated by decreased expression of α‐smooth muscle antigen and vimentin and preserved expression of E‐cadherin in the CG‐injured peritoneum and cultured human peritoneal mesothelial cells exposed to transforming growth factor‐β1. Nintedanib also suppressed expression of Snail and Twist, two transcription factors associated with MMT in vivo and in vitro. Moreover, nintedanib treatment inhibited expression of several cytokines/chemokines, including tumour necrosis factor‐α, interleukin‐1β and interleukin‐6, monocyte chemoattractant protein‐1 and prevented infiltration of macrophages to the injured peritoneum. Finally, nintedanib reduced CG‐induced peritoneal vascularization. These data suggest that nintedanib may attenuate peritoneal fibrosis by inhibiting MMT, inflammation, and angiogenesis and have therapeutic potential for the prevention and treatment of peritoneal fibrosis in patients on peritoneal dialysis.

Delayed Administration of Nintedanib Ameliorates Fibrosis Progression in CG-Induced Peritoneal Fibrosis Mouse Model

“…Means with different superscript symbols (* ,#,@ ) are significantly different from one another. partner, Smad3, was able to activate VEGFR and Src to stimulate peritoneal angiogenesis [9]. Inflammation plays an essential role in the development of peritoneal fibrosis, including monocytes/macrophages filtration, proinflammatory cytokines production by inflammatory cells, fibroblasts, and mesenchymal cells via EMT from PMCs, and this process can promote ECM protein synthesis, EMT, angiogenesis, and inflammation [29].…”

“…The peritoneal fibrosis model was established in male C57/BL6 mouse that weighed 24–28 g (Shanghai Super–B&K Laboratory Animal Corp. Ltd.), as described in our previous study [ 9 ]. Briefly, peritoneal fibrosis in mice was generated by intraperitoneal injection of 0.1% CG dissolved in 0.9% saline every other day for 21 days or 35 days.…”

“…Formalin-fixed samples of peritoneum were embedded in paraffin and processed to prepare 3-μm-thick sections. Immunohistochemical staining was conducted as described in our previous study [ 9 ]. To evaluate peritoneal fibrosis, Masson trichrome staining was performed according to the protocol provided by the manufacturer (Sigma-Aldrich).…”

“…Immunoblot analysis of peritoneum tissue samples was conducted as described previously [ 9 ]. The densitometry analysis of immunoblot results was conducted using Image J software developed at the National Institute of Health.…”

“…Because of its anti-fibrotic properties, nintedanib was one of the first drugs approved for use in idiopathic pulmonary fibrosis (IPF) and has recently further been approved for treatment of progressive fibrosing interstitial lung diseases (ILDs) and systemic sclerosis-associated ILD [ 2 , 3 ]. Inspired by this breakthrough, many researchers, including our team, have examined the anti-fibrotic effect of nintedanib in other tissues and organs and found its anti-fibrotic effects in the liver, skin, kidney, and peritoneum [ 4 , 5 , 6 , 7 , 8 , 9 ].…”