Selective expression of mitogen-inducible cyclooxygenase in macrophages stimulated with lipopolysaccharide

Soo H. Lee, Emmanuel Soyoola, Prithiva Chanmugam, Suzanne Hart, Wenqin Sun, Hua Zhong, Shuenn Liou, Daniel Simmons, Daniel Hwang

Research output: Contribution to journalArticle

742 Scopus citations

Abstract

Two forms of cyclooxygenase are known to be present in eukaryotic organisms: a cyclooxygenase (COX-1) first purified from ram seminal vesicles encoded by a 2.8-kilobase mRNA, and a newly discovered mitogen-inducible cyclooxygenase (COX-2) encoded by a 4-kilobase mRNA. Expression of these two forms of the enzyme in rat alveolar macrophages stimulated with lipopolysaccharide was investigated by 1) determining the activity of newly synthesized enzyme after inactivating the endogenous enzyme with aspirin; 2) comparing levels of newly synthesized enzyme proteins in cells treated with or without lipopolysaccharide; and 3) assessing the expression of the mRNAs encoding COX-1 and COX-2. Levels of enzyme proteins were assessed by Western blot analysis and immunoprecipitation of 35S-labeled enzyme using two different antibodies, one specific for COX-2 and the other recognizing both forms of the enzyme but preferentially recognizing COX-1. We report here that the enhanced cyclooxygenase activity induced by the bacterial lipopolysaccharide in rat alveolar macrophages is caused by selective expression of the COX-2. Expression of COX-2 in macrophages stimulated by lipopolysaccharide was completely inhibited by dexamethasone, whereas COX-1 was unaffected. In resting unstimulated macrophages, only COX-1 but not COX-2 was detected. Levels of mRNA for the COX-2 in macrophages were increased, but those of the COX-1 were not affected by lipopolysaccharide as assessed by reverse transcription coupled with polymerase chain reaction. These results indicate that increased synthesis of prostaglandins and thromboxanes in lipopolysaccharide-stimulated macrophages results from selective expression of COX-2.

Original languageEnglish (US)
Pages (from-to)25934-25938
Number of pages5
JournalJournal of Biological Chemistry
Volume267
Issue number36
StatePublished - Dec 25 1992
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Selective expression of mitogen-inducible cyclooxygenase in macrophages stimulated with lipopolysaccharide'. Together they form a unique fingerprint.

  • Cite this

    Lee, S. H., Soyoola, E., Chanmugam, P., Hart, S., Sun, W., Zhong, H., Liou, S., Simmons, D., & Hwang, D. (1992). Selective expression of mitogen-inducible cyclooxygenase in macrophages stimulated with lipopolysaccharide. Journal of Biological Chemistry, 267(36), 25934-25938.