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Modulation of tissue plasminogen activator and plasminogen activator inhibitor-1 by transforming growth factor-beta in human retinal glial cells.
Invest Ophthalmol Vis Sci 2002; 43(8):2799-805IO

Abstract

PURPOSE

The serine proteases tissue plasminogen activator (t-PA) and urokinase plasminogen activator (u-PA) and their inhibitor, plasminogen activator inhibitor (PAI)-1, regulate a variety of processes involved in tissue morphogenesis and differentiation. There is much evidence that plasminogen activator-mediated extracellular matrix degradation is an important step in the development of ocular neovascular diseases. The authors investigated whether expression of t-PA, u-PA, and PAI-1 in human retinal glial cells (HRGCs) is influenced by exposure to transforming growth factor (TGF)-beta, a cytokine that regulates the proliferation and differentiation of cells.

METHODS

The extracellular release of t-PA, u-PA, and PAI-1 was measured by enzyme-linked immunosorbent assay (ELISA) in the supernatant of HRGC cultures, under basal conditions and after stimulation with TGF-beta at various concentrations (2, 5, 10, or 20 ng/mL). Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze mRNA levels. Smad2 phosphorylation was detected by Western blot analysis.

RESULTS

Under basal conditions, HRGCs secreted considerable amounts of t-PA and PAI-1. Stimulation with TGF-beta resulted in increased synthesis of t-PA and PAI-1 protein in a time- and dose-dependent manner. Moreover, an increased expression of t-PA and PAI-1 mRNA after supplementation with TGF-beta was observed, with maximum expression at 12 hours. In contrast, HRGCs did not respond to TGF-beta with any change of u-PA production, although there were detectable amounts of u-PA mRNA and protein. Phosphorylation of Smad2 was increased after addition of TGF-beta. This effect was partially reversible after treatment with interferon-gamma.

CONCLUSIONS

The production of plasminogen activators and PAI-1 by HRGCs reflects the potential role of these cells in the progression of neovascular ocular diseases. Furthermore, the finding that t-PA and PAI-1 synthesis by HRGCs is mediated by TGF-beta and its downstream effector Smad2 confirms the importance of the TGF-beta signaling pathway in the regulation of interactions between retinal cells and the extracellular matrix.

Authors+Show Affiliations

Department of Ophthalmology, Johann Wolfgang Goethe University Hospital, Frankfurt am Main, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12147618

Citation

Schacke, Wolfgang, et al. "Modulation of Tissue Plasminogen Activator and Plasminogen Activator Inhibitor-1 By Transforming Growth Factor-beta in Human Retinal Glial Cells." Investigative Ophthalmology & Visual Science, vol. 43, no. 8, 2002, pp. 2799-805.
Schacke W, Beck KF, Pfeilschifter J, et al. Modulation of tissue plasminogen activator and plasminogen activator inhibitor-1 by transforming growth factor-beta in human retinal glial cells. Invest Ophthalmol Vis Sci. 2002;43(8):2799-805.
Schacke, W., Beck, K. F., Pfeilschifter, J., Koch, F., & Hattenbach, L. O. (2002). Modulation of tissue plasminogen activator and plasminogen activator inhibitor-1 by transforming growth factor-beta in human retinal glial cells. Investigative Ophthalmology & Visual Science, 43(8), pp. 2799-805.
Schacke W, et al. Modulation of Tissue Plasminogen Activator and Plasminogen Activator Inhibitor-1 By Transforming Growth Factor-beta in Human Retinal Glial Cells. Invest Ophthalmol Vis Sci. 2002;43(8):2799-805. PubMed PMID: 12147618.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Modulation of tissue plasminogen activator and plasminogen activator inhibitor-1 by transforming growth factor-beta in human retinal glial cells. AU - Schacke,Wolfgang, AU - Beck,Karl-Friedrich, AU - Pfeilschifter,Josef, AU - Koch,Frank, AU - Hattenbach,Lars-Olof, PY - 2002/7/31/pubmed PY - 2002/8/8/medline PY - 2002/7/31/entrez SP - 2799 EP - 805 JF - Investigative ophthalmology & visual science JO - Invest. Ophthalmol. Vis. Sci. VL - 43 IS - 8 N2 - PURPOSE: The serine proteases tissue plasminogen activator (t-PA) and urokinase plasminogen activator (u-PA) and their inhibitor, plasminogen activator inhibitor (PAI)-1, regulate a variety of processes involved in tissue morphogenesis and differentiation. There is much evidence that plasminogen activator-mediated extracellular matrix degradation is an important step in the development of ocular neovascular diseases. The authors investigated whether expression of t-PA, u-PA, and PAI-1 in human retinal glial cells (HRGCs) is influenced by exposure to transforming growth factor (TGF)-beta, a cytokine that regulates the proliferation and differentiation of cells. METHODS: The extracellular release of t-PA, u-PA, and PAI-1 was measured by enzyme-linked immunosorbent assay (ELISA) in the supernatant of HRGC cultures, under basal conditions and after stimulation with TGF-beta at various concentrations (2, 5, 10, or 20 ng/mL). Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze mRNA levels. Smad2 phosphorylation was detected by Western blot analysis. RESULTS: Under basal conditions, HRGCs secreted considerable amounts of t-PA and PAI-1. Stimulation with TGF-beta resulted in increased synthesis of t-PA and PAI-1 protein in a time- and dose-dependent manner. Moreover, an increased expression of t-PA and PAI-1 mRNA after supplementation with TGF-beta was observed, with maximum expression at 12 hours. In contrast, HRGCs did not respond to TGF-beta with any change of u-PA production, although there were detectable amounts of u-PA mRNA and protein. Phosphorylation of Smad2 was increased after addition of TGF-beta. This effect was partially reversible after treatment with interferon-gamma. CONCLUSIONS: The production of plasminogen activators and PAI-1 by HRGCs reflects the potential role of these cells in the progression of neovascular ocular diseases. Furthermore, the finding that t-PA and PAI-1 synthesis by HRGCs is mediated by TGF-beta and its downstream effector Smad2 confirms the importance of the TGF-beta signaling pathway in the regulation of interactions between retinal cells and the extracellular matrix. SN - 0146-0404 UR - https://www.unboundmedicine.com/medline/citation/12147618/Modulation_of_tissue_plasminogen_activator_and_plasminogen_activator_inhibitor_1_by_transforming_growth_factor_beta_in_human_retinal_glial_cells_ L2 - http://iovs.arvojournals.org/article.aspx?volume=43&page=2799 DB - PRIME DP - Unbound Medicine ER -