Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts

Mohan R K Dasu, Robert E. Barrow, Marcus Spies, David Herndon

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

In this study, we investigated the effect of inflammatory cytokines on matrix metalloproteinase (MMP-1) and TIMP-1 production in human dermal fibroblasts, which play a pivotal role in wound healing, ranging from the synthesis and remodeling of extracellular matrix (ECM) to the synthesis of growth factors. The balance of MMPs and TIMPs is crucial in directing successful wound repair. Human adult dermal fibroblasts were seeded in six well plates (7.5×104 cells/ml) in complete media. Eighty to ninety percent confluent cells were treated with interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (10ng/ml) for 6h in serum free media with suitable controls run in triplicate. Supernatants were assayed for pro-MMP-1 & TIMP-1. Extracted total RNA was used for reverse transcription polymerase chain reaction (RT-PCR) with sequence specific primers for MMP-1, TIMP-1 and β-actin. Signal intensity was normalized to the internal control (β-actin). Statistical analysis used ANOVA. MMP-1 and TIMP-1 mRNA expression were markedly increased with IL-6 and TNF-α treatment and remains unchanged with IL-1β. Pro-MMP-1 protein levels are unchanged with TNF-α and significantly increased with IL-1β and IL-6 treatment. However, TNF-α significantly increases TIMP-1 protein levels. Data suggests differential regulation of MMP-1 and TIMP-1 protein levels by the cytokines found in stimulated dermal fibroblasts. Further characterization of this response will provide an understanding of the mechanisms of pathogenesis of extracellular matrix (ECM) and the potential role of metalloproteinases in tissue remodeling after injury.

Original languageEnglish (US)
Pages (from-to)527-531
Number of pages5
JournalBurns
Volume29
Issue number6
DOIs
StatePublished - Sep 2003

Fingerprint

Tissue Inhibitor of Metalloproteinase-1
Matrix Metalloproteinases
Fibroblasts
Cytokines
Skin
Tumor Necrosis Factor-alpha
Interleukin-1
Tissue Inhibitor of Metalloproteinases
Interleukin-6
Extracellular Matrix
Actins
Matrix Metalloproteinase 1
Serum-Free Culture Media
Wounds and Injuries
Metalloproteases
Wound Healing
Reverse Transcription
Intercellular Signaling Peptides and Proteins
Analysis of Variance
RNA

Keywords

  • Cytokines
  • Dermal fibroblasts
  • Extracellular matrix
  • MMP-1
  • TIMP-1

ASJC Scopus subject areas

  • Emergency Medicine
  • Surgery

Cite this

Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts. / Dasu, Mohan R K; Barrow, Robert E.; Spies, Marcus; Herndon, David.

In: Burns, Vol. 29, No. 6, 09.2003, p. 527-531.

Research output: Contribution to journalArticle

Dasu, Mohan R K ; Barrow, Robert E. ; Spies, Marcus ; Herndon, David. / Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts. In: Burns. 2003 ; Vol. 29, No. 6. pp. 527-531.
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