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Title: Nitric oxide and the activity of NADPH oxidase in rat granulocytes after stimulation with lipopolysaccharide in vitro


In neutrophils the respiratory burst depend on the activation of NADPH oxidase. We stimulated neutrophils with LPS for 3 h and then the respiratory burst in vitro was examined by luminol dependent chemiluminescence (CL). LPS did not cause an increased respiratory burst as compared to control cells. Activation of cells by PMA induced a robust CL in neutrophils stimulated with LPS. Neutrophils stimulated with LPS and activated by PMA demonstrated more than a threefold increase of the respiratory burst. TIRON and apocynin significantly decreased the respiratory burst of neutrophils, indicating that the main source of •O in activated neutrophils was NADPH oxidase. Sodium azide had no impact on the respiratory burst of neutrophils. Selective inhibitors of iNOS: L-NIL and 1400W caused reversion of the values of respiratory burst in cells stimulated with LPS to the values obtained for control cells. This finding suggests that nitric oxide participates in the respiratory burst. Production of NO by LPS-stimulated cells was completely abolished by the iNOS inhibitors and actinomycin D. This result indicates that iNOS is the major source of NO in LPS-stimulated neutrophils. Stimulated cells showed a rapid induction of iNOS mRNA and functional protein. Activation of neutrophils in the presence of SNAP showed that NO (within a narrow concentration range) enhances the respiratory burst of ne ; utrophils. This process is inversely proportional to the concentration of SNAP.This thesis demonstrates that in rat neutrophils administration of LPS induces a rapid expression of iNOS and stimulates the production of NO. Then, nitric oxide affects the activity of constitutive NADPH oxidase, leading to increased production of •O .

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Level of degree:

2 - studia doktoranckie

Degree discipline:

farmakologia ; biochemia

Degree grantor:

Wydział Lekarski


Uracz, Wojciech

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tylko w bibliotece

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Last modified:

Mar 17, 2023

In our library since:

Mar 6, 2013

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Edition name Date
ZB-116230 Mar 17, 2023


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