Matrix Metalloproteinase Inhibitors Cause Cell Cycle Arrest and Apoptosis in Glomerular Mesangial Cells
- Catherine Daniel1,
- Jeremy Duffield2,
- Thomas Brunner3,
- Karin Steinmann-Niggli1,
- Nadège Lods1 and
- Hans-Peter Marti1
- 1Division of Nephrology and Hypertension, University of Bern, Bern, Switzerland (C.D., K.S.-N., N.L., H.-P.M.); 2Center for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom (J.D.); and 3Institute of Pathology, University of Bern, Bern, Switzerland (T.B.)
Abstract
Inflammation is characterized by an excess of cell proliferation often leading to fibrosis and sclerosis with subsequent loss of organ function. We hypothesized that these features may be ameliorated by induction of cell cycle arrest and apoptosis as result of therapy with matrix metalloproteinase (MMP) inhibitors. In our study, mesangial cells and experimental mesangial proliferative glomerulonephritis provided the model of inflammation. First, we investigated the effect of the MMP inhibitor BB-1101 in anti-Thy1.1 nephritis. The numbers of apoptotic glomerular cells in nephritic rats increased about 4 and 6 times as a result of BB-1101 therapy, observed 11 and 14 days after induction of disease, respectively. Subsequently, rat mesangial cells were exposed to an MMP inhibitor in vitro. Fluorescence-activated cell sorter analyses of cells exposed to RO111-3456 demonstrated a dose-dependent cell cycle arrest in the G0/G1phase associated with increased expression of statin. The cell cycle arrest was followed by apoptosis as investigated by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) biotin nick-end labeling (TUNEL) and acridine orange/ethidium bromide stainings, as well as by annexin V binding. The induction of p53, p21, and bax, but not the Fas/FasL pathway appeared to play an important pathogenetic role. In summary, MMP inhibitors induce cell cycle arrest followed by apoptosis in mesangial cells. These features help to explain the anti-inflammatory effects of these compounds, such as reduction of mesangial cell proliferation and attenuation of extracellular matrix accumulation. In conclusion, induction of cell cycle arrest with subsequent apoptosis may offer new perspectives in the therapy of inflammation even beyond kidney diseases.
Footnotes
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Send reprint requests to: Hans-Peter Marti, M.D., Division of Nephrology and Hypertension, Inselspital Bern, CH-3010 Bern, Switzerland. E-mail: hmarti{at}insel.ch
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This work was supported by Grants 31-49765.96 and 31-55779.98 to H.-P.M. from the Swiss National Foundation for Scientific Research. Portions of the study were presented as poster at the 32nd Annual Meeting of the American Society of Nephrology, November 5–8, 1999, Miami Beach, FL. Daniel C, Duffield J, Thomas G, Ziswiler RA, Steinmann-Niggli K and Marti HP (1999) Matrix metalloproteinase inhibitor induces apoptosis in mesangial cells. J Am Soc Nephrol10:569A.
- Abbreviations:
- MMP
- matrix metalloproteinase
- ECM
- extracellular matrix
- PBS
- phosphate-buffered saline
- DMSO
- dimethyl sulfoxide
- TUNEL
- terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) biotin nick-end labeling
- FCS
- fetal calf serum
- LDH
- lactate dehydrogenase
- PAGE
- polyacrylamide gel electrophoresis
- AO/EB
- acridine orange/ethidium bromide
- FACS
- fluorescence-activated cell sorter
- PI
- propidium iodide
- TBS-T
- Tris-buffered saline-Tween 20
- DEVD-CHO
- acetyl-Asp-Glu-Val-Asp-aldehyde
- FITC
- fluorescein isothiocyanate
- TNF-α
- tumor necrosis factor-α
- IETD-AFC
- N-acetyl-Ile-Glu-Thr-Asp-7-amino-4-trifluoromethyl coumarin
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- Received September 26, 2000.
- Accepted December 19, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
