Upregulation of Transglutaminase andε(γ-Glutamyl)-Lysine in the Fisher-Lewis Rat Model of Chronic Allograft Nephropathy

Abstract

Background. Tissue transglutaminase (TG2), a cross-linking enzyme, modulates deposition of extracellular matrix protein in renal fibrosis. This study aimed to examine TG2 and its cross-link productε(γ-glutamyl)-lysine in the Fisher-Lewis rat renal transplantation (RTx) model of chronic allograft nephropathy (CAN).Materials and Methods. Left renal grafts from male Fisher and Lewis were transplanted into Lewis rats, generating allografts and isografts, respectively. Blood pressure, renal function, and proteinuria were monitored for up to 52 weeks. At termination, CAN was assessed in the renal tissue by light and electron microscopy, TG2 andε(γ-glutamyl)-lysine by immunofluorescence, and the urinaryε(γ-glutamyl)-lysine by high performance liquid chromatography.Results. Compared to the isograft, the allografts were hypertensive, proteinuric, and uraemic and developed CAN. Extracellular TG2 (glomerulus:64.55±17.61versus2.11±0.17,P<0.001; interstitium:13.72±1.62versus3.19±0.44,P<0.001),ε(γ-glutamyl)-lysine (glomerulus:21.74±2.71versus1.98±0.37,P<0.01; interstitium:37.96±17.06versus0.42±0.11,P<0.05), TG2 enzyme activity (1.09±0.13versus0.41±0.03 nmol/h/mg protein,P<0.05), TG2 mRNA (20-fold rise), and urinaryε(γ-glutamyl)-lysine (534.2±198.4 nmol/24 h versus57.2±4.1 nmol/24 h,P<0.05) levels were significantly elevated in the allografts and showed a positive linear correlation with tubulointerstitial fibrosis.Conclusion. CAN was associated with upregulation of renal TG2 pathway, which has a potential for pharmacological intervention. The elevated urinaryε(γ-glutamyl)-lysine, measured for the first time in RTx, is a potential biomarker of CAN.