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Am J Physiol Renal Physiol 274: F1113-F1118, 1998;
0363-6127/98 $5.00
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Vol. 274, Issue 6, F1113-F1118, June 1998

Estradiol reverses TGF-beta 1-stimulated type IV collagen gene transcription in murine mesangial cells

Sharon Silbiger1, Jun Lei1, Fuad N. Ziyadeh2, and Joel Neugarten1

1 Nephrology Division, Department of Medicine, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10467; and 2 Penn Center for Molecular Studies of Kidney Diseases, Renal-Electrolyte and Hypertension Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

We have previously shown that estradiol suppresses types I and IV collagen synthesis by mesangial cells grown in the presence of serum. In the present study, we examined the interaction between estradiol and transforming growth factor-beta (TGF-beta ) on collagen IV synthesis. In a luciferase reporter gene construct containing the type IV collagen promoter and alpha 1-chain regulatory sequences, we found that TGF-beta 1 (2 ng/ml) stimulated alpha 1-collagen IV gene transcription in serum-free media (140.5 ± 6.2 relative luciferase units, expressed as a percent of control untreated cells, P < 0.001). Estradiol reversed the stimulatory effects of TGF-beta 1 on reporter gene transcription in a dose-dependent manner [for 2.5 × 10-9 M, 114.2 ± 0.2, P < 0.002 vs. TGF-beta 1; for 10-7 M, 89.5 ± 4.0, P < 0.001 vs. TGF-beta 1 and P = not significant (NS) vs. control]. Using immunoprecipitation techniques, we found that estradiol (10-7 M) reversed TGF-beta 1-stimulated type IV collagen synthesis (175.3 ± 14.7 vs. 111.6 ± 7.1, expressed as a percent of control untreated cells, P < 0.001) but did not affect TGF-beta 1-stimulated type I collagen synthesis (166.9 ± 18.8 vs. 162.2 ± 16.2, P = NS). These results were confirmed with Western blotting. Nuclear extracts from mesangial cells treated with TGF-beta 1 showed increased binding to a Sp1 consensus binding sequence oligonucleotide and to an Sp1 binding site in the collagen IV promoter. Estradiol reversed this enhanced binding. These data suggest that estradiol antagonizes TGF-beta 1-stimulated type IV collagen synthesis at a transcriptional level and that this effect may be mediated by interactions with the transcription factor Sp1.

mesangial cells; transforming growth factor-beta ; Sp1


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