Divergent regulation of 92-kDa gelatinase and TIMP-1 by HBECs in response to IL-1beta and TNF-alpha

Am J Physiol. 1997 Oct;273(4):L866-74. doi: 10.1152/ajplung.1997.273.4.L866.

Abstract

In this study, we addressed the question of whether human bronchial epithelial cells (HBECs) contribute to the regulation of 92-kDa gelatinase activity by secreting tissue inhibitor of metalloproteinase (TIMP)-1. We investigated expression of 92-kDa gelatinase and TIMP-1 in response to lipopolysaccharide (LPS) and to the proinflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha. Confluent HBECs from explants were cultured in plastic dishes coated with type I and III collagen. We demonstrated that TIMP-1 was expressed at both the protein and mRNA levels by primary cultures of HBECs. Gelatin zymography of HBEC-conditioned media showed that exposure of HBECs to LPS, IL-1beta, or TNF-alpha induced a twofold increase in the latent form of 92-kDa gelatinase production, as well as its activation. Also, quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR) demonstrated a twofold increase in the 92-kDa mRNA level in response to both cytokines. In contrast, TIMP-1 production evaluated by immunoblotting was unchanged in the presence of LPS and IL-1beta and was clearly decreased in the presence of TNF-alpha. Quantitative RT-PCR demonstrated that TIMP-1 mRNA levels remained unchanged in response to LPS or IL-1beta but decreased by 70% in the presence of TNF-alpha. All of these results strongly suggest that the control mechanisms regulating the expression of 92-kDa gelatinase and TIMP-1 by HBECs in response to inflammatory stimuli are divergent and result in an imbalance between 92-kDa gelatinase and TIMP-1 in favor of the metalloproteinase. Such an imbalance may contribute significantly to acute airway inflammation.

MeSH terms

  • Breast
  • Bronchi / metabolism*
  • Cells, Cultured
  • Culture Media, Conditioned
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Female
  • Fibroblasts / metabolism
  • Gelatinases / biosynthesis*
  • Gene Expression Regulation / drug effects*
  • Humans
  • Interleukin-1 / pharmacology*
  • Kinetics
  • Lipopolysaccharides / pharmacology
  • Macrophages, Alveolar / metabolism
  • Macrophages, Alveolar / pathology
  • Molecular Weight
  • Polymerase Chain Reaction
  • RNA, Messenger / biosynthesis
  • Respiratory Distress Syndrome / metabolism
  • Respiratory Distress Syndrome / pathology
  • Tissue Inhibitor of Metalloproteinase-1 / biosynthesis*
  • Transcription, Genetic / drug effects
  • Tumor Necrosis Factor-alpha / pharmacology*

Substances

  • Culture Media, Conditioned
  • Interleukin-1
  • Lipopolysaccharides
  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinase-1
  • Tumor Necrosis Factor-alpha
  • Gelatinases