Increased Legumain/Smad3 expression in atherosclerotic plaque of rat thoracic aorta

Biomed Pharmacother. 2019 Nov:119:109353. doi: 10.1016/j.biopha.2019.109353. Epub 2019 Sep 12.

Abstract

Objectives: The purpose of this study was to investigate the role of legumain in the formation and stability of atherosclerotic plaque, as well as to explore the association between legumain with Smad3 pathway in a rat atherosclerosis model.

Methods: Rat with thoracic aorta atherosclerosis was established and received treatment with statin (n = 15 each) or controls (n = 10). Serum level of legumain was determined by enzyme-linked immunosorbent assay. Legumain and Smad3 aortic expression levels were assessed by immunohistochemistry and fluorescence microscopy. Protein and mRNA levels were analyzed using Western blot analysis and reverse transcriptase coupled polymerase chain reaction, respectively.

Results: The atherosclerotic group showed higher serum legumain level than control and statin group. Expression of legumain and Smad3 in macrophages and foam cells was increased in atherosclerotic group compared to control and statin group. The protein and mRNA levels of legumain and Smad3 were significantly attenuated by statin treatment (p < 0.05). For all groups, legumain expression was correlated linearly with Smad3 at mRNA (coefficient: 0.94) and protein (coefficient: 097) level.

Conclusions: Legumain and Smad3 expression is highly expressed in mainly atherosclerotic plaque macrophages and linearly related, which is attenuated by statin therapy, suggesting legumain a potential Smad3 pathway-related marker of atherosclerosis.

Keywords: Atherosclerosis; Inflammatory response; Legumain; Macrophages; Smad3.

MeSH terms

  • Animals
  • Aorta, Thoracic / metabolism*
  • Aorta, Thoracic / pathology*
  • Body Weight
  • Cysteine Endopeptidases / blood
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / metabolism*
  • Lipids / blood
  • Male
  • Plaque, Atherosclerotic / blood
  • Plaque, Atherosclerotic / genetics
  • Plaque, Atherosclerotic / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats, Wistar
  • Smad3 Protein / genetics
  • Smad3 Protein / metabolism*

Substances

  • Lipids
  • RNA, Messenger
  • Smad3 Protein
  • Cysteine Endopeptidases
  • asparaginylendopeptidase