Nephrol Dial Transplant (2001) 16: 1776-1782
© 2001 European Renal Association-European Dialysis and Transplant Association
Cultured rat glomerular epithelial cells show gene expression and production of transforming growth factor-ß: expression is enhanced by thrombin
Second Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
Background. Glomerular crescents play an important role in progressive glomerular injury. The lesions consist of epithelial cells, macrophages, and deposits of fibrin and extracellular matrix. Transforming growth factor beta (TGF-ß) contributes to the modulation of cell growth and extracellular matrix synthesis. Thrombin is involved in fibrin formation in crescents. The purpose of this study was to examine whether glomerular epithelial cells (GEC) could produce TGF-ß, and if so, to clarify the role of TGF-ß in GEC proliferation. We also investigated whether thrombin could modulate the production of TGF-ß and extracellular matrix by GEC.
Methods. Bioassay using the TGF-ß-dependent mink pulmonary epithelial cell line (CCL-64), immunoblot analysis, and reverse transcriptase polymerase chain reaction (RT-PCR) were used to demonstrate TGF-ß production by rat GEC. TGF-ß gene expression was examined by RT-PCR in GEC incubated with thrombin, and type IV collagen and fibronectin were quantified by enzyme immunoassay in culture supernatants of GEC incubated with thrombin or TGF-ß.
Results. TGF-ß activity was demonstrated in GEC supernatants by bioassay. Immunoblot analysis of concentrated culture supernatants using anti-TGF-ß antibody revealed a 12.5-kDa protein, which was compatible with TGF-ß. Concentrated GEC supernatants inhibited GEC proliferation as well as porcine TGF-ß. RT-PCR demonstrated TGF-ß gene expression in GEC. Thrombin (0.5–5.0 U/ml) enhanced TGF-ß mRNA expression in a dose-dependent manner. Thrombin (5.0 U/ml) and porcine TGF-ß (5.0 ng/ml) stimulated the production of type IV collagen and fibronectin by GEC.
Conclusions. Rat GEC produce TGF-ß in vitro. Thrombin may participate in the progression of glomerulosclerosis in crescentic glomerulonephritis through the stimulation of TGF-ß production by GEC.
Keywords: fibronectin; glomerular epithelial cells; rat; TGF-ß; thrombin; type IV collagen
Correspondence and offprint requests to: Hideaki Yamabe MD, Second Department of Internal Medicine, Hirosaki University School of Medicine, Zaifu-cho 5, Hirosaki 036-8216, Japan.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  T. Fujita, H. Yamabe, M. Shimada, R. Murakami, R. Kumasaka, N. Nakamura, H. Osawa, and K. Okumura Thrombin enhances the production of monocyte chemoattractant protein-1 and macrophage inflammatory protein-2 in cultured rat glomerular epithelial cells Nephrol. Dial. Transplant., November 1, 2008; 23(11): 3412 - 3417. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. C.J. Howell, R. H. Johns, J. A. Lasky, B. Shan, C. J. Scotton, G. J. Laurent, and R. C. Chambers Absence of Proteinase-Activated Receptor-1 Signaling Affords Protection from Bleomycin-Induced Lung Inflammation and Fibrosis Am. J. Pathol., May 1, 2005; 166(5): 1353 - 1365. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Shirato, H. Osawa, M. Kaizuka, N. Nakamura, T. Sugawara, M. Nakamura, M. Tamura, H. Yamabe, and K. Okumura Thrombin stimulates production of fibronectin by human proximal tubular epithelial cells via a transforming growth factor-{beta}-dependent mechanism Nephrol. Dial. Transplant., November 1, 2003; 18(11): 2248 - 2254. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Pavenstadt, W. Kriz, and M. Kretzler Cell Biology of the Glomerular Podocyte Physiol Rev, January 1, 2003; 83(1): 253 - 307. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Ding, K. Reddy, A. A. Kapasi, N. Franki, N. Gibbons, B. S. Kasinath, and P. C. Singhal Angiotensin II induces apoptosis in rat glomerular epithelial cells Am J Physiol Renal Physiol, July 1, 2002; 283(1): F173 - F180. [Abstract] [Full Text] [PDF]
|
|