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Glomerular epithelial cells secrete a glomerular basement membrane-degrading metalloproteinase
R Johnson1, H Yamabe, Y P Chen
1Department of medicine, University of Washington Medical Center, Seattle.
Journal of the American Society of Nephrology : JASN
|March 1, 1992
Summary
Cultured rat glomerular epithelial cells (GEC) release metalloproteinases that degrade extracellular matrix. These enzymes may play a key role in glomerular basement membrane remodeling and kidney diseases.
Area of Science:
- Nephrology
- Cell Biology
- Biochemistry
Background:
- Glomerular epithelial cells (GEC) are crucial for kidney function.
- Understanding GEC's role in extracellular matrix (ECM) remodeling is vital for kidney disease research.
Purpose of the Study:
- To investigate the capacity of cultured rat GEC to release ECM-degrading proteinases.
- To characterize the activity and substrate specificity of these proteinases.
Main Methods:
- Cultured rat GEC were used to collect conditioned media.
- Gelatinase activity was measured using [3H]gelatin as a substrate.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with gelatin or type IV collagen substrates was employed.
- Northern blot analysis was performed to identify metalloproteinases.
Main Results:
- GEC-conditioned media exhibited gelatinase activity, primarily at neutral pH.
- Several proteinase bands were detected, with a major band at 150 kDa.
- The identified metalloproteinases were distinct from those found in mesangial cells or neutrophils.
- GEC proteinases effectively degraded type IV collagen from the glomerular basement membrane.
Conclusions:
- Cultured rat GEC produce multiple neutral metalloproteinases.
- These GEC-derived metalloproteinases degrade components of the glomerular basement membrane.
- GEC metalloproteinases may be significant in glomerular basement membrane remodeling and associated kidney diseases.