Effect of glucose on the expression of type I collagen and transforming growth factor-beta1 in cultured human peritoneal fibroblasts.
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)-beta1 in human peritoneal fibroblasts in culture.


METHODS
Prospective experimental study.


METHODS
University medical center.


METHODS
Primary cultures of fibroblasts established from peritoneal tissues of five patients. High glucose treatment of the primary cultured fibroblasts.


METHODS
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 beta-actin with TGF-beta1 or type I collagen mRNAs was used to quantitate type I collagen and TGF-beta1 mRNA levels in response to increasing glucose concentrations with and without TGF-beta1 antibody treatment.


RESULTS
There was a significant increase in the mRNA for type I collagen and TGF-beta1 in response to increasing glucose concentrations in a dose response-dependent manner. The TGF-beta1 antibody treatment resulted in an 83% and 68% decrease in type I collagen and TGF-beta1 mRNA levels, respectively.


CONCLUSIONS
Increasing glucose concentrations stimulated type I collagen expression in human peritoneal fibroblasts in culture. A potential mediator for this effect is TGF-beta1. 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.
Saed
Ghassan M
GM
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, and the C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan 48201, USA. gsaed@wayne.edu
Diamond
Michael P
MP
D000906:Antibodies / Q000494:pharmacology; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics*; D018076:DNA, Complementary; D003850:Deoxyribonuclease I; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D010537:Peritoneum / Q000166:cytology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000276:immunology; D053773:Transforming Growth Factor beta1
D000906:Antibodies; D024042:Collagen Type I; D018076:DNA, Complementary; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D003850:Deoxyribonuclease I; D005947:Glucose