Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo

Kirkwood A. Pritchard, M. Kerry O'Banion, Joseph M. Miano, Niksa Vlasic, Umesh G. Bhatia, Donald A. Young, Michael B. Stemerman

Research output: Contribution to journalArticle

Abstract

Prostaglandins are synthesized from arachidonic acid by the rate-limiting enzyme cyclooxygenase (prostaglandin G/H synthase). Active cyclooxygenase is encoded by two distinct and independently regulated genes, termed cyclooxygenase-1 (cox1) and cyclooxygenase-2 (cox2). In this investigation, we examined the expression of cox1 and cox2 mRNA in rat aorta following balloon deendothelialization (BDE) in vivo and in rat aortic smooth muscle cells (SMC) after serum stimulation in vitro. Two h after BDE, rat aortic cox2 mRNA levels increased greater than 50-fold relative to the lowest detectable levels on days 2 and 14. No message was detectable in non-BDE control rat aortas. Similar to the results found in vivo, cultured SMC exhibited a greater than 45-fold increase in cox2 mRNA levels after a 2-h exposure to serum. This increase was transient because cox2 levels declined at 4 and 8 h. In contrast, minimal changes in cox1 mRNA levels were observed after BDE or serum treatments. Increased levels of cox2 mRNA and corresponding protein synthesis led to an accumulation of total cyclooxygenase protein, which remained elevated 24 h after serum stimulation. Serum-treated SMC also generated greater amounts of cyclooxygenase-dependent metabolites than quiescent SMC as evidenced by marked increases in prostaglandin E2 content in conditioned media. This increase is associated with a 2.5-3.0-fold increased rate of arachidonic acid conversion to prostaglandin E2. Our data indicate that injury and serum stimulation differentially regulate mRNA and protein expression of two distinct cox genes in vascular SMC in vivo and in vitro. The findings suggest that the prostanoid responses after vascular injury are, in part, mediated by acute increases in cox2 mRNA and cyclooxygenase-2 protein.

Original languageEnglish (US)
Pages (from-to)8504-8509
Number of pages6
JournalJournal of Biological Chemistry
Volume269
Issue number11
StatePublished - Mar 18 1994
Externally publishedYes

Fingerprint

Cyclooxygenase 2
Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Rats
Cells
Prostaglandin-Endoperoxide Synthases
Messenger RNA
Cyclooxygenase 1
Balloons
Serum
Dinoprostone
Arachidonic Acid
Prostaglandins
Aorta
Proteins
Genes
Rat control
In Vitro Techniques
Vascular System Injuries

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Pritchard, K. A., O'Banion, M. K., Miano, J. M., Vlasic, N., Bhatia, U. G., Young, D. A., & Stemerman, M. B. (1994). Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. Journal of Biological Chemistry, 269(11), 8504-8509.

Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. / Pritchard, Kirkwood A.; O'Banion, M. Kerry; Miano, Joseph M.; Vlasic, Niksa; Bhatia, Umesh G.; Young, Donald A.; Stemerman, Michael B.

In: Journal of Biological Chemistry, Vol. 269, No. 11, 18.03.1994, p. 8504-8509.

Research output: Contribution to journalArticle

Pritchard, KA, O'Banion, MK, Miano, JM, Vlasic, N, Bhatia, UG, Young, DA & Stemerman, MB 1994, 'Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo', Journal of Biological Chemistry, vol. 269, no. 11, pp. 8504-8509.
Pritchard KA, O'Banion MK, Miano JM, Vlasic N, Bhatia UG, Young DA et al. Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. Journal of Biological Chemistry. 1994 Mar 18;269(11):8504-8509.
Pritchard, Kirkwood A. ; O'Banion, M. Kerry ; Miano, Joseph M. ; Vlasic, Niksa ; Bhatia, Umesh G. ; Young, Donald A. ; Stemerman, Michael B. / Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 11. pp. 8504-8509.
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abstract = "Prostaglandins are synthesized from arachidonic acid by the rate-limiting enzyme cyclooxygenase (prostaglandin G/H synthase). Active cyclooxygenase is encoded by two distinct and independently regulated genes, termed cyclooxygenase-1 (cox1) and cyclooxygenase-2 (cox2). In this investigation, we examined the expression of cox1 and cox2 mRNA in rat aorta following balloon deendothelialization (BDE) in vivo and in rat aortic smooth muscle cells (SMC) after serum stimulation in vitro. Two h after BDE, rat aortic cox2 mRNA levels increased greater than 50-fold relative to the lowest detectable levels on days 2 and 14. No message was detectable in non-BDE control rat aortas. Similar to the results found in vivo, cultured SMC exhibited a greater than 45-fold increase in cox2 mRNA levels after a 2-h exposure to serum. This increase was transient because cox2 levels declined at 4 and 8 h. In contrast, minimal changes in cox1 mRNA levels were observed after BDE or serum treatments. Increased levels of cox2 mRNA and corresponding protein synthesis led to an accumulation of total cyclooxygenase protein, which remained elevated 24 h after serum stimulation. Serum-treated SMC also generated greater amounts of cyclooxygenase-dependent metabolites than quiescent SMC as evidenced by marked increases in prostaglandin E2 content in conditioned media. This increase is associated with a 2.5-3.0-fold increased rate of arachidonic acid conversion to prostaglandin E2. Our data indicate that injury and serum stimulation differentially regulate mRNA and protein expression of two distinct cox genes in vascular SMC in vivo and in vitro. The findings suggest that the prostanoid responses after vascular injury are, in part, mediated by acute increases in cox2 mRNA and cyclooxygenase-2 protein.",
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