Pharmacological modulation of transglutaminase 2 in the unilateral ureteral obstruction mouse model

Eur J Pharmacol. 2024 Dec 5:984:177037. doi: 10.1016/j.ejphar.2024.177037. Epub 2024 Oct 5.

Abstract

Background: Transglutaminase 2 (TG2) is a multifunctional enzyme involved in fibrosis by promoting transforming-growth-factor-β1 and crosslinking of extracellular matrix proteins. These functions are dependent on the open conformation, while the closed state of TG2 can induce vasodilation. We explored the putative protective role of TG2 in its closed state on development of renal fibrosis and blood pressure (BP) regulation.

Methods: We studied the unilateral ureteral obstruction (UUO) mouse model treated with LDN27219, which promotes the closed conformation of TG2. Mice were subjected to 7 days UUO or sham operation and treated with vehicle (n = 10), LDN27219 (15 mg/kg/12 h, n = 9) or candesartan (5 mg/kg/day, n = 10) as a clinically comparator. Renal expression of TG2 and pro-fibrotic mediators were evaluated by Western blotting, qPCR and histology, and BP by tail-cuff measurements.

Results: Obstructed kidneys showed increased mRNA and protein expression of fibronectin, collagen 3α1 (Col3α1), α-smooth muscle actin and collagen staining. Despite increased renal TG2 mRNA, protein expression was reduced in all UUO groups, but with increased transamidase activity in the vehicle and candesartan groups. LDN27219 reduced mRNA expression of fibronectin and Col3α1, but their protein expression remained unchanged. In contrast to LDN27219, candesartan lowered BP without affecting expression of pro-fibrotic biomarkers.

Conclusion: Renal TG2 mRNA and protein expression levels seem dissociated, with transamidase activity being increased. LDN27219 influences kidney pro-fibrotic markers at the mRNA level and attenuates transamidase activity but without affecting collagen content or BP. Our findings suggest that TG2 in its closed conformation has anti-fibrotic effects at the molecular level.

Keywords: Fibrosis; Kidney disease; Pharmacology; Transglutaminase 2; Unilateral ureteral obstruction model.

MeSH terms

  • Actins / genetics
  • Actins / metabolism
  • Animals
  • Benzimidazoles / pharmacology
  • Benzimidazoles / therapeutic use
  • Biphenyl Compounds / pharmacology
  • Biphenyl Compounds / therapeutic use
  • Blood Pressure / drug effects
  • Collagen Type III / genetics
  • Collagen Type III / metabolism
  • Disease Models, Animal*
  • Fibronectins / genetics
  • Fibronectins / metabolism
  • Fibrosis*
  • GTP-Binding Proteins* / genetics
  • GTP-Binding Proteins* / metabolism
  • Kidney* / drug effects
  • Kidney* / metabolism
  • Kidney* / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Protein Glutamine gamma Glutamyltransferase 2*
  • Tetrazoles / pharmacology
  • Transglutaminases* / genetics
  • Transglutaminases* / metabolism
  • Ureteral Obstruction* / complications
  • Ureteral Obstruction* / drug therapy
  • Ureteral Obstruction* / pathology

Substances

  • Transglutaminases
  • Protein Glutamine gamma Glutamyltransferase 2
  • GTP-Binding Proteins
  • Tetrazoles
  • Biphenyl Compounds
  • candesartan
  • Collagen Type III
  • Benzimidazoles
  • Fibronectins
  • Actins
  • COL3A1 protein, mouse