Tubulointerstitial injury leading to
fibrosis is a common pathway of many renal diseases. During this type of injury, modeled by unilateral
ureteral obstruction (UUO), cells undergo epithelial-to-mesenchymal transition (EMT), a process that is mediated by various
cytokines that modulate the biology of
extracellular matrix proteins. Here, we studied the
tubulointerstitial nephritis antigen (TINag), a tubular basement membrane
protein, in the UUO model of tubulointerstitial injury. We observed upregulation of
type IV collagen but downregulation of both
laminin and TINag in obstructed kidneys. TINag downregulation was a result of oxidative stress; in the proximal tubular epithelial cell line HK-2, TINag expression and its promoter activity decreased
after treatment with H2O2. We identified multiple
CCAAT/enhancer binding protein beta (
C/EBP-beta) motifs in the TINag promoter and observed that
oxidant stress perturbed interactions between TINag
DNA and
C/EBP-beta protein.
Oxidant stress reduced nuclear translocation of
C/EBP-beta in HK-2 cells, which was restored by
antioxidants. In addition, overexpression of
C/EBP-beta restored the H2O2-induced reduction of TINag promoter activity and expression. Furthermore, in vivo, renal obstruction reduced nuclear expression of
C/EBP-beta. Cells grown on a TINag substratum maintained their normal epithelial phenotype and cytoskeletal organization, similar to those grown on
type IV collagen, and demonstrated reduced synthesis of
fibronectin. Taken together, these findings suggest that altered interactions between
C/EBP-beta and TINag play a critical role in the pathophysiology of renal injury after obstruction.