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Hypoxia and cytokines differentially regulate the expression of the tissue plasminogen activator (tPA) and plasminogen activator inhibitor‐1 (PAI‐1) genes in cultured human mesothelial cells (HMCs): Implications for post‐surgical peritoneal fibrinolytic dysregulation
Author(s) -
Keenan Benjamin J,
Currie Brian M,
Heydrick Stanley J,
Stucchi Arthur F
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.717.5
Subject(s) - t plasminogen activator , plasminogen activator , hypoxia (environmental) , plasminogen activator inhibitor 1 , tissue plasminogen activator , medicine , endocrinology , fibrinolysis , messenger rna , cytokine , inflammation , chemistry , biology , gene , biochemistry , oxygen , organic chemistry
Abdominal surgery induces mesothelial hypoxia and inflammation, both implicated in the dysregulation of the peritoneal fibrinolytic system and intraabdominal adhesion formation. Our objective examined the contribution of each using cultured HMCs. Cells were treated with either a cytokine (C) cocktail (TGFβ+IL‐1β+IFN‐γ), the hypoxia mimetic dimethyloxaloylglycine (D) or both, and the expression of tPA and PAI‐1 mRNA was measured at 1, 3, 6, 12, and 18 hours (n=4/group) by RT‐PCR. C alone and C+D increased PAI‐1 mRNA levels at 6 hr (2.4±0.1 fold; p<0.001 and 2.1±0.17 fold; p<0.001 vs controls) and 12 hr (4.1±0.4 fold; p<0.001 and 4.1±0.7 fold; p<0.001 vs controls) while D alone elevated PAI‐1 mRNA levels at 6 and 12 hr (p<0.05). D increased tPA mRNA levels at 12 hr (2.2±0.16 fold; p<0.001) and 18 hr (3.2±0.7 fold; p<0.001) vs control while C+D paralleled closely at both time points (p<0.001). The tPA/PAI‐1 mRNA ratio, a predictor of fibrinolytic capacity, was reduced 40% on average (p<0.001) at 6 hr in all groups vs control. However, it rose thereafter in D and C+D (2.2±0.4 fold; p<0.001 and 3.8±0.7 fold; p<0.001 vs 6 hr) while C continued to drop at 12 hr. Our results show that cytokines appear to regulate PAI‐1 while hypoxia regulates tPA, suggesting that both contribute to peritoneal fibrinolytic dysregulation and adhesiogenesis. Funded by Cooper‐Tyler Endowment, Department of Surgery, Boston University Medical Center.