Effect of glucose on the expression of type I collagen and transforming growth factor-β1 in cultured human peritoneal fibroblasts

Ghassan M. Saed, Michael Peter Diamond

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Objective: To determine the effect of high glucose levels on the expression of type I collagen, and whether this effect is regulated by transforming growth factor (TGF)-β1 in human peritoneal fibroblasts in culture. Design: Prospective experimental study. Setting: University medical center. Patient(s): Primary cultures of fibroblasts established from peritoneal tissues of five patients. Intervention(s): High glucose treatment of the primary cultured fibroblasts. Main Outcome Measure(s): Primary cultures of human peritoneal fibroblasts were incubated with varying amounts of glucose (1-5 g/L) for 24 hours. Total RNA was extracted from human peritoneal fibroblasts and converted to cDNA by reverse transcriptase. Multiplex reverse transcriptase/polymerase chain reaction (PCR) simultaneously co-amplifying β-actin with TGF-β1 or type I collagen mRNAs was used to quantitate type I collagen and TGF-β1 mRNA levels in response to increasing glucose concentrations with and without TGF-β1 antibody treatment. Result(s): There was a significant increase in the mRNA for type I collagen and TGF-β1 in response to increasing glucose concentrations in a dose response-dependent manner. The TGF-β1 antibody treatment resulted in an 83% and 68% decrease in type I collagen and TGF-β1 mRNA levels, respectively. Conclusion(s): Increasing glucose concentrations stimulated type I collagen expression in human peritoneal fibroblasts in culture. A potential mediator for this effect is TGF-β1. These results have implications not only for individuals with diabetes mellitus who may be predisposed to greater postoperative adhesion development, but also for individuals with surgical stress responses after surgery.

Original languageEnglish (US)
Pages (from-to)158-163
Number of pages6
JournalFertility and sterility
Volume79
Issue number1
DOIs
StatePublished - Jan 1 2003

Fingerprint

Transforming Growth Factors
Collagen Type I
Fibroblasts
Glucose
Messenger RNA
Antibodies
Multiplex Polymerase Chain Reaction
RNA-Directed DNA Polymerase
Reverse Transcriptase Polymerase Chain Reaction
Actins
Diabetes Mellitus
Research Design
Therapeutics
Complementary DNA
Outcome Assessment (Health Care)
Prospective Studies
RNA

Keywords

  • Clinical study
  • Fibroblasts
  • Postoperative adhesions
  • RT/PCR
  • TGF-β1
  • Type I collagen

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology

Cite this

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title = "Effect of glucose on the expression of type I collagen and transforming growth factor-β1 in cultured human peritoneal fibroblasts",
abstract = "Objective: To determine the effect of high glucose levels on the expression of type I collagen, and whether this effect is regulated by transforming growth factor (TGF)-β1 in human peritoneal fibroblasts in culture. Design: Prospective experimental study. Setting: University medical center. Patient(s): Primary cultures of fibroblasts established from peritoneal tissues of five patients. Intervention(s): High glucose treatment of the primary cultured fibroblasts. Main Outcome Measure(s): Primary cultures of human peritoneal fibroblasts were incubated with varying amounts of glucose (1-5 g/L) for 24 hours. Total RNA was extracted from human peritoneal fibroblasts and converted to cDNA by reverse transcriptase. Multiplex reverse transcriptase/polymerase chain reaction (PCR) simultaneously co-amplifying β-actin with TGF-β1 or type I collagen mRNAs was used to quantitate type I collagen and TGF-β1 mRNA levels in response to increasing glucose concentrations with and without TGF-β1 antibody treatment. Result(s): There was a significant increase in the mRNA for type I collagen and TGF-β1 in response to increasing glucose concentrations in a dose response-dependent manner. The TGF-β1 antibody treatment resulted in an 83{\%} and 68{\%} decrease in type I collagen and TGF-β1 mRNA levels, respectively. Conclusion(s): Increasing glucose concentrations stimulated type I collagen expression in human peritoneal fibroblasts in culture. A potential mediator for this effect is TGF-β1. These results have implications not only for individuals with diabetes mellitus who may be predisposed to greater postoperative adhesion development, but also for individuals with surgical stress responses after surgery.",
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N2 - Objective: To determine the effect of high glucose levels on the expression of type I collagen, and whether this effect is regulated by transforming growth factor (TGF)-β1 in human peritoneal fibroblasts in culture. Design: Prospective experimental study. Setting: University medical center. Patient(s): Primary cultures of fibroblasts established from peritoneal tissues of five patients. Intervention(s): High glucose treatment of the primary cultured fibroblasts. Main Outcome Measure(s): Primary cultures of human peritoneal fibroblasts were incubated with varying amounts of glucose (1-5 g/L) for 24 hours. Total RNA was extracted from human peritoneal fibroblasts and converted to cDNA by reverse transcriptase. Multiplex reverse transcriptase/polymerase chain reaction (PCR) simultaneously co-amplifying β-actin with TGF-β1 or type I collagen mRNAs was used to quantitate type I collagen and TGF-β1 mRNA levels in response to increasing glucose concentrations with and without TGF-β1 antibody treatment. Result(s): There was a significant increase in the mRNA for type I collagen and TGF-β1 in response to increasing glucose concentrations in a dose response-dependent manner. The TGF-β1 antibody treatment resulted in an 83% and 68% decrease in type I collagen and TGF-β1 mRNA levels, respectively. Conclusion(s): Increasing glucose concentrations stimulated type I collagen expression in human peritoneal fibroblasts in culture. A potential mediator for this effect is TGF-β1. These results have implications not only for individuals with diabetes mellitus who may be predisposed to greater postoperative adhesion development, but also for individuals with surgical stress responses after surgery.

AB - Objective: To determine the effect of high glucose levels on the expression of type I collagen, and whether this effect is regulated by transforming growth factor (TGF)-β1 in human peritoneal fibroblasts in culture. Design: Prospective experimental study. Setting: University medical center. Patient(s): Primary cultures of fibroblasts established from peritoneal tissues of five patients. Intervention(s): High glucose treatment of the primary cultured fibroblasts. Main Outcome Measure(s): Primary cultures of human peritoneal fibroblasts were incubated with varying amounts of glucose (1-5 g/L) for 24 hours. Total RNA was extracted from human peritoneal fibroblasts and converted to cDNA by reverse transcriptase. Multiplex reverse transcriptase/polymerase chain reaction (PCR) simultaneously co-amplifying β-actin with TGF-β1 or type I collagen mRNAs was used to quantitate type I collagen and TGF-β1 mRNA levels in response to increasing glucose concentrations with and without TGF-β1 antibody treatment. Result(s): There was a significant increase in the mRNA for type I collagen and TGF-β1 in response to increasing glucose concentrations in a dose response-dependent manner. The TGF-β1 antibody treatment resulted in an 83% and 68% decrease in type I collagen and TGF-β1 mRNA levels, respectively. Conclusion(s): Increasing glucose concentrations stimulated type I collagen expression in human peritoneal fibroblasts in culture. A potential mediator for this effect is TGF-β1. These results have implications not only for individuals with diabetes mellitus who may be predisposed to greater postoperative adhesion development, but also for individuals with surgical stress responses after surgery.

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