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Cyclooxygenase-2 is expressed in human fibroblasts isolated from intraperitoneal adhesions but not from normal
Ghassan M Saed1, Adnan R Munkarah, Michael P Diamond
1Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Detroit, Michigan 48201, USA. g.saed@wayne.edu
Objective:
To determine whether the COX-2 gene is expressed in human fibroblasts isolated from normal peritoneal and adhesion tissues.
Design:
Prospective experimental study.
Setting:
University medical center.
Patient(S):
Five patients undergoing laparotomy for pelvic pain. Primary cultures of fibroblasts were taken from both peritoneum and adhesion tissues.
Intervention(S):
Hypoxia treatment of the primary cultured fibroblasts.
Main Outcome Measures:
We used the multiplex reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry techniques to determine whether COX-2 mRNA and its protein were present in normal peritoneal and adhesion fibroblasts from the same patients. Total RNA was extracted from cultured fibroblasts and subjected to multiplex RT-PCR to detect the presence of COX-2 mRNA in these cells. Cultured fibroblasts from all tissues were also fixed on slides and stained with COX-2 monoclonal antibody labeled with immunofluorescence.
Result(S):
COX-2 mRNA and its protein were absent in normal peritoneal fibroblasts from all five subjects but were present in adhesion fibroblasts from the same patients, as indicated by the multiplex RT-PCR and immunohistochemistry techniques. Hypoxia treatment significantly induced the mRNA and COX-2 protein levels in normal peritoneal fibroblasts to levels seen in adhesion fibroblasts under normoxic conditions. However, hypoxia had no effects on COX-2 expression by adhesion fibroblasts.
Conclusion(S):
Adhesion fibroblasts develop a specific phenotype, an adhesion phenotype, which is in part characterized by the expression of COX-2. The expression of COX-2 mRNA in adhesion fibroblasts and the induction of COX-2 in peritoneal fibroblasts in response to hypoxia indicate a possible inflammatory response. Regulation of COX-2 may alter peritoneal healing and may provide the opportunity to reduce postoperative adhesion development.