Effect of the p38 MAP kinase inhibitor SB203580 on cytokine-mediated priming of arachidonate release in neutrophils
| Stimulus, 1 μM . | SB203580, μM . | Priming agent, % control . | ||
|---|---|---|---|---|
| FCS . | TNF-α . | GM-CSF . | ||
| FMLP | 0 | — | 100+ | 100 |
| 1 | — | 136 ± 36 | 125 ± 11 | |
| 5 | — | 113 ± 27 | 159 ± 24 | |
| 10 | — | 107 ± 26 | 145 ± 18 | |
| 15 | — | 107 ± 23 | 149 ± 19 | |
| 20 | — | 109 ± 33 | 130 ± 19 | |
| 30 | — | 91 ± 20 | 78 ± 30 | |
| A23187 | 0 | 100 | 100 | 100 |
| 1 | 74 ± 13 | 187 ± 12* | 130 ± 12 | |
| 5 | 91 ± 12 | 199 ± 19† | 138 ± 16 | |
| 10 | 76 ± 23 | 211 ± 14* | ND | |
| 15 | 80 ± 28 | 216 ± 27† | 115 ± 8 | |
| 20 | 97 ± 21 | 206 ± 27† | 115 ± 9 | |
| 30 | 112 ± 31 | 182 ± 22† | 127 ± 11 | |
| Stimulus, 1 μM . | SB203580, μM . | Priming agent, % control . | ||
|---|---|---|---|---|
| FCS . | TNF-α . | GM-CSF . | ||
| FMLP | 0 | — | 100+ | 100 |
| 1 | — | 136 ± 36 | 125 ± 11 | |
| 5 | — | 113 ± 27 | 159 ± 24 | |
| 10 | — | 107 ± 26 | 145 ± 18 | |
| 15 | — | 107 ± 23 | 149 ± 19 | |
| 20 | — | 109 ± 33 | 130 ± 19 | |
| 30 | — | 91 ± 20 | 78 ± 30 | |
| A23187 | 0 | 100 | 100 | 100 |
| 1 | 74 ± 13 | 187 ± 12* | 130 ± 12 | |
| 5 | 91 ± 12 | 199 ± 19† | 138 ± 16 | |
| 10 | 76 ± 23 | 211 ± 14* | ND | |
| 15 | 80 ± 28 | 216 ± 27† | 115 ± 8 | |
| 20 | 97 ± 21 | 206 ± 27† | 115 ± 9 | |
| 30 | 112 ± 31 | 182 ± 22† | 127 ± 11 | |
Purified neutrophils were incubated with SB203580 for 30 minutes at 37°C before priming with either 500 U/mL TNF-α, 10 ng/mL GM-CSF, or 0.01% FCS diluent for 20 minutes followed by stimulation with agonist as indicated for 15 minutes. Arachidonate release was measured as described in “Materials and methods.” Basal release of arachidonate in unstimulated samples was subtracted from the stimulated value, and the data were expressed as a percentage of the DMSO diluent control. Absolute values expressed as cpm/106 cells for (1) FMLP stimulation: FCS, 118 ± 100; TNF, 1493 ± 461; and GM-CSF, 1317 ± 321 and (2) A23187 stimulation: FCS, 2584 ± 786; TNF, 4856 ± 1094; and GM-CSF, 11 306 ± 3332. In all cases the release of arachidonate from unprimed FMLP-stimulated cells was not significantly different from background. The data shown are the mean ± SE of 4 experiments.
ND indicates not determined; FCS, fetal calf serum; TNF-α, tumor necrosis factor–α; GM-CSF, granulocyte-macrophage colony-stimulating factor; FMLP, formyl-methionyl-leucyl-phenylalanine.
.01 > P > .001 indicates significant difference between samples with inhibitor.
.05 > P > .01 indicates significant difference between samples without inhibitor.