Increased oxidative metabolism in PMA-activated lymphocytes: A flow cytometric study

Rabesandratana, Herisoa; Fournier, Anne-Marie; Château, Marie-Thérèse; Serre, A; Dornand, Jacques

doi:10.1016/0192-0561(92)90089-4

Cited by 38 publications

(8 citation statements)

References 20 publications

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“…There are several controversies in studies concerning the effect of the addition of SOD on oxidative stress in cells as monitored with DCFH. Some studies reported an inhibitory effect by SOD [20–22], whereas almost the same number of studies reported that SOD had no effect [23–25]. The finding made here, that SOD inhibited the oxidation of DCFH, could provide an explanation for discrepancies regarding the effect of SOD on the DCFH test for oxidative stress.…”

Section: Discussionmentioning

confidence: 50%

Rapid oxidation of dichlorodihydrofluorescin with heme and hemoproteins: formation of the fluorescein is independent of the generation of reactive oxygen species

et al. 2001

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Oxidative stress and the generation of reactive oxygen species (ROS) have been implicated in the pathogenesis of cellular damage. These events have usually been reported in terms of oxidation of a reporter molecule such as 2P P,7P Pdichlorodihydrofluorescin diacetate (DCFH-DA). Treatment of HeLa cells with hemin or metalloporphyrins resulted in a rapid oxidation of DCFH in a time-and dose-dependent manner. This oxidation was inhibited by treatment of the cells with a large amount of superoxide dismutase and catalase, which is different from observations that these enzymes had no effect on the induction of heme oxygenase-1, a stress-induced protein, in hemin-treated cells. To examine the possibility that the oxidation of DCFH is independent of the generation of ROS, the oxidation was measured using hemoglobin-synthesizing erythroleukemia K562 cells. When K562 cells were treated with N N-aminolevulinic acid, a precursor of heme, oxidation of DCFH increased depending on the heme content in cells. Then DCFH-DA was oxidized directly with heme, hemoglobin, myoglobin and cytochrome c. These results suggest that oxidation of DCFH is not always related to the generation of ROS but may be related to heme content in cells. ß

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Section: Discussionmentioning

confidence: 50%

Rapid oxidation of dichlorodihydrofluorescin with heme and hemoproteins: formation of the fluorescein is independent of the generation of reactive oxygen species

et al. 2001

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“…PMA (Rabesandratana et al , 1992; Demiryurek et al , 1994; Lundqvist and Dahlgren, 1996) and PAF (Takahashi et al , 1991; Kato et al , 2002) have been well documented to stimulate the production of ROS from leucocytes. In the present study, both PMA and PAF produced concentration‐dependent increases in ROS generation in PAR‐2+/+ mice lymphocytes, which were not modified by PAR‐2 deficiency as shown by their responses in lymphocytes from PAR‐2−/− mice.…”

Section: Discussionmentioning

confidence: 99%

Activation of mouse protease‐activated receptor‐2 induces lymphocyte adhesion and generation of reactive oxygen species

Lim

Tennant

Kennedy

et al. 2006

British J Pharmacology

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Background and Purpose: Protease-activated receptor-2 (PAR-2) is expressed on lymphocytes and endothelial cells, and plays a significant role in inflammatory reactions. Since leukocyte-endothelial cell interaction and reactive oxygen species (ROS) generation are hallmarks of the development of inflammation, the effects of PAR-2 activation by trypsin on lymphocyte adhesion and ROS generation was examined utilising PAR-2 wild type and knockout (PAR-2À/À) mice. Experimental Approach: Lymphocyte adhesion to the luminal surface of mouse isolated aortae was measured using 51 Crlabelled leukocytes and ROS generation from isolated lymphocytes was quantified using chemiluminescence. Key results: Trypsin induced adhesion of lymphocytes when added exogenously to the endothelial surface of the aorta for 30 min. Similarly, increased lymphocyte adhesion was also observed when mice were injected with trypsin intravenously 24 h prior to the adhesion assay, an effect which was partly ICAM-1 mediated. Trypsin also increased ROS generation from isolated mouse lymphocytes in a dose-dependent manner. The increase in lymphocyte adhesion and ROS production in response to trypsin were abolished in PAR-2À/À mice indicating a PAR-2 dependent mechanism. Superoxide dismutase had a greater inhibitory effect in PAR-2À/À mice compared to wild type mice when lymphocytes were stimulated with PMA but not trypsin. Conclusions and Implications: The present study indicates that activation of PAR-2 may be an important factor in modulating lymphocyte adhesion and ROS generation. The results have implications for developing anti-inflammatory strategies.

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“…Intracellular oxidative metabolism was assayed using dichlorofluorescein diacetate (DCFH-DA) as reported [23]. Dichlorofluorescein diacetate is a cell-permeable molecule that is cleaved to non-fluorescent DCF that remains trapped in the cytoplasmic compartment.…”

Section: Measurement Of Reactive Intracellular Oxidative Metabolites mentioning

confidence: 99%

Inhibition of fMLP-triggered respiratory burst of human monocytes by adenosine: involvement of A3 adenosine receptor

Broussas

Cornillet‐Lefèbvre

Potron

et al. 1999

Journal of Leukocyte Biology

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Increased oxidative metabolism in PMA-activated lymphocytes: A flow cytometric study

Cited by 38 publications

References 20 publications

Rapid oxidation of dichlorodihydrofluorescin with heme and hemoproteins: formation of the fluorescein is independent of the generation of reactive oxygen species

Rapid oxidation of dichlorodihydrofluorescin with heme and hemoproteins: formation of the fluorescein is independent of the generation of reactive oxygen species

Activation of mouse protease‐activated receptor‐2 induces lymphocyte adhesion and generation of reactive oxygen species

Inhibition of fMLP-triggered respiratory burst of human monocytes by adenosine: involvement of A3 adenosine receptor

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