A Single Competitive Reverse Transcriptase-Polymerase Chain Reaction Assay to Measure Inducible Nitric Oxide Synthase Gene Expression in Both Human and Rat

1997 ◽  
Vol 247 (2) ◽  
pp. 455-458 ◽  
Author(s):  
Gong Chen ◽  
Timothy D. Warner
Keyword(s):  
Gene Expression ◽  
Nitric Oxide ◽  
Polymerase Chain Reaction ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Polymerase Chain Reaction Assay ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Effect of NOS Inhibitor on Cytokine and COX2 Expression in Rat Pulpitis

2005 ◽  
Vol 84 (8) ◽  
pp. 762-767 ◽  
Author(s):  
N. Kawashima ◽  
H. Nakano-Kawanishi ◽  
N. Suzuki ◽  
M. Takagi ◽  
H. Suda
Keyword(s):  
Nitric Oxide ◽  
Polymerase Chain Reaction ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Reverse Transcription ◽  
Chain Reaction ◽  
Nos Inhibitor ◽  
Polymerase Chain ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Various kinds of chemical mediators are synthesized in the course of pulpitis; thus, control of their production would assist in inducing a reduction in pulpal inflammation. We hypothesized that nitric oxide (NO) would be an important mediator of pulpal inflammation. Pulpal inflammation was induced by the application of LPS in rat incisor pulp, and inducible nitric oxide synthase (iNOS) expression was evaluated by reverse-transcription/polymerase chain-reaction and immunohistochemical staining. After LPS application, iNOS mRNA was first detected after 3 hrs, peaked at 6 hrs, and decreased thereafter. iNOS-positive cells were macrophages and neutrophils. An NOS inhibitor caused drastic decreases in the expression of pro-inflammatory cytokines and COX2 mRNA, which was highly induced in the LPS-induced pulpitis. These results indicate that NO synthesis is related to the initiation of mediator production, and that its down-regulation should contribute to the prevention of pro-inflammatory mediator synthesis. Abbreviations: ANOVA, analysis of variance; COX2, cyclo-oxygenase 2; EDTA, ethylenediaminetetraacetic acid; iNOS, inducible nitric oxide synthase; IL, interleukin; L-NAME, NG-nitro L-arginine methyl ester; LPS, lipopolysaccharide; NO, nitric oxide; NOS, nitric oxide synthases; PG, prostaglandin; RT-PCR, reverse-transcription/polymerase chain-reaction; TNFα, tumor necrosis factor alpha.

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