Mesangial cell gelatinase A synthesis is attenuated by oscillating hyperbaric pressure

Biochem J. 2002 Mar 15;362(Pt 3):693-700. doi: 10.1042/0264-6021:3620693.

Abstract

Glomerular hypertension has been established as a major factor contributing to glomerular scarring. Underlying cellular mechanisms leading to matrix accumulation are largely unknown. The isolated effect of oscillating hyperbaric pressure [OP; P(max) 50 mmHg (1 mmHg=0.133 kPa), P(mean) 24 mmHg, with a fixed oscillation of 60/min] on matrix-degrading protease secretion by rat mesangial cells (MCs) was analysed using a pressure chamber model described previously [Mertens, Espenkott, Venjakob, Heintz, Handt and Sieberth (1998) Hypertension 32, 945-952]. MCs were grown under atmospheric pressure (AP) or a controlled OP, and protease synthesis and gene transcription were analysed. A distinct biphasic cellular response to OP with stimulated gelatinase A protein expression and enzyme activity during the initial 24 h, and subsequent inhibition, was apparent, as shown by gelatin zymography. Gelatinase B activity remained unchanged. The abundance of gelatinase A transcripts, determined by reverse transcriptase-PCR, indicated a concordant regulation of gene transcription. To elucidate underlying regulatory events, reporter constructs were transfected. In these experiments, a recently identified response element, RE-1, conferred a significant stimulatory effect within the initial 4 h of OP. Nuclear protein/RE-1 binding studies revealed additional complexes from 5 min up to 3 h after OP exposure, with intensities dependent on P(max). STAT3 was identified as a component of these novel complexes. Down-regulation of cis-activity after 48 h of OP exposure was not transferred via the proximal 1686 bp of the gelatinase A regulatory sequence. In conclusion, hyperbaric OP elicits time-dependent changes in rat MC gelatinase A gene transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Nucleus / enzymology
  • Cells, Cultured
  • DNA Primers
  • Gene Expression Regulation, Enzymologic
  • Glomerular Mesangium / enzymology*
  • Hypertension, Renal / enzymology
  • Kinetics
  • Matrix Metalloproteinase 2 / genetics*
  • Matrix Metalloproteinase Inhibitors
  • Oscillometry
  • Polymerase Chain Reaction
  • Pressure*
  • Rats
  • Time Factors
  • Transcription, Genetic*

Substances

  • DNA Primers
  • Matrix Metalloproteinase Inhibitors
  • Matrix Metalloproteinase 2