Table 1.
Effect of misoprostol treatment before or after lipopolysaccharide (LPS) stimulation on equine leukocyte extracellular cytokine levels.
| Cytokine | Misoprostol (μM) |
||
|---|---|---|---|
| 0 µM | 100 µM Pretreatment | 100 µM Posttreatment | |
| (A) Analyte levels 6 h post LPS treatment (pg/mL) | |||
| Tumor necrosis factor α (TNFα) | 894.064 (±179.221)* | 106.758 (±18.060)† | 123.098 (±30.886)† |
| IL-6 | 261.540 (±42.707)* | 189.514 (±27.817)† | 142.422 (±26.252)† |
| IL-1β | 3,150.524 (±1,520.336)§ | 6,027.748 (±2,914.889)‡ | 3,594.648 (±1,682.129) |
| IL-8 | 160.750 (±33.802)* | 158.802 (±30.795) | 154.986 (±30.643) |
| IFNγ-induced protein 10 (IP-10) | 173.014 (±51.100)* | 168.298 (±52.347) | 166.196 (±50.940)† |
| IL-10 | 284.534 (±59.435)* | 322.008 (±79.808) | 269.366 (±53.206) |
| IL-4 | 1,486.826 (±557.649)* | 1,468.370 (±550.651) | 1,460.358 (±543.439) |
| Interferon γ (IFNγ) | 947.748 (±286.373)* | 932.092 (±281.452) | 918.024 (±285.689) |
| IL-2 | 12.988 (±2.803)* | 12.984 (±2.635) | 12.988 (±2.687) |
| IL-18 | 142.850 (±43.023)* | 143.136 (±42.121) | 142.644 (±42.195) |
| IL-5 | 56.785 (±14.970)* | 58.262 (±14.692) | 56.848 (±14.935) |
| Fractalkine | 1,211.792 (±772.502)* | 1,210.620 (±795.108) | 1,203.280 (±776.168) |
| Granulocyte-stimulating factor (G-CSF) | 334.736 (±100.147)* | 337.046 (±99.541) | 329.058 (±101.121) |
| GRO | 206.730 (±19.548)* | 217.976 (±19.254) | 210.944 (±19.566) |
| Monocyte chemotactic protein-1 (MCP-1) | 3,767.000 (±1,012.198)* | 3,770.200 (±1,004.467) | 3,686.400 (±990.298) |
| Eotaxin | 15.282 (±6.499) | 15.618 (±6.714) | 15.008 (±6.685) |
| (B) Analyte levels 24 h post LPS treatment (pg/mL) | |||
| TNFα | 159.388 (±32.838)* | 27.758 (±3.707)† | 35.814 (±9.906)† |
| IL-6 | 93.298 (±14.794)* | 59.310 (±12.267)† | 52.858 (±10.413)† |
| IL-1β | 1,133.266 (±366.477)§ | 670.146 (±214.096) | 575.286 (±200.162)‡ |
| IL-8 | 67.866 (±10.138)* | 68.048 (±9.256) | 67.058 (±8.910) |
| IP-10 | 90.624 (±19.337) | 82.244 (±23.320) | 84.344 (±21.908) |
| IL-10 | 163.348 (±38.802)* | 153.022 (±33.184) | 147.664 (±35.273) |
| IL-4 | 389.620 (±121.471) | 393.586 (±124.957) | 395.160 (±125.627) |
| IFNγ | 617.860 (±202.735) | 283.524 (±67.384) | 251.630 (±72.228) |
| IL-2 | 5.688 (±0.769) | 5.694 (±0.677) | 5.550 (±0.791) |
| IL-18 | 35.224 (±8.172) | 35.786 (±8.486) | 35.934 (±8.32) |
| IL-5 | 16.934 (±3.445) | 17.280 (±3.481) | 17.166 (±3.444) |
| Fractalkine | 2.294 (±0.233) | 2.314 (±0.232) | 2.306 (±0.243) |
| G-CSF | 85.532 (±28.732) | 89.294 (±29.413) | 89.166 (±31.782) |
| GRO | 62.574 (±7.753)* | 61.424 (±6.904) | 63.848 (±6.862) |
| MCP-1 | 778.114 (217.657) | 797.044 (±215.033) | 791.620 (±214.179) |
| Eotaxin | – | – | – |
Four different treatment groups were analyzed: unstimulated equine leukocyte-rich plasma (LRP) pretreated with the misoprostol vehicle, 0.05% DMSO (baseline values; not shown in table).
0 µM = equine LRP pretreated for 30 min with misoprostol vehicle (0.05% DMSO) before LPS stimulation; 100 µM Pretreatment = equine LRP pretreated for 30 min with 100 µM misoprostol before LPS stimulation; 100 µM Posttreatment = equine LRP was stimulated with 100 ng/mL LPS, followed 30 min later by treatment with 100 µM misoprostol.
All samples were incubated at 37°C for (A) 6 or (B) 24 h. Cell supernatants were analyzed in triplicate for cytokine/chemokine levels using a multiplex bead-based immunoassay. Data are presented as mean ± SEM cytokine levels in pg/mL. Significant differences from unstimulated samples pretreated with the misoprostol vehicle (not shown) via one-way RM ANOVA (*p < 0.05) or Friedman RM ANOVA (§p < 0.05) are indicated. Significant differences from LPS-stimulated samples pretreated with the misoprostol vehicle (denoted as 0 µM misoprostol) via one-way RM ANOVA (†p < 0.05) or Friedman RM ANOVA on Ranks (‡p < 0.05) are also indicated; n = 5.