HaCaT Eicosanoid Analysis Results
The aim of this study was to investigate the levels of AA derived lipid mediators in the supernatant following stimulation of cells with A23187 (with and without COX inhibition), ATP and UVR. LC-MS/MS was used for the eicosanoid analysis and the values were normalized using cell numbers (pg/10^6 cells).
Calcium Ionophore
Time (h) | Mediator | |||||
---|---|---|---|---|---|---|
PGE2 | 15-keto-PGE2 | 13,14-dihydro-15-keto-PGE2 | PGF2α | 12-HETE | 15-HETE | |
0 | 9.45 ± 7.56 | 0.50 ± 0.00 | 2.00 ± 0.00 | 7.00 ± 0.00 | 6.44 ± 4.40 | 3.72 ± 1.08 |
0.5 | 601.26 ± 162.02 | 65.52 ± 57.95 | 211.38 ± 61.05 | 106.06 ± 39.89 | 19.49 ± 10.36 | 163.26 ± 143.10 |
1 | 14371.34 ± 858.76 | 2136.87 ± 588.92 | 2461.84 ± 452.79 | 2787.86 ± 489.04 | 363.25 ± 96.91 | 4050.14 ± 2998.16 |
3 | 79576.88 ± 15355.19 | 12230.83 ± 2179.66 | 11025.93 ± 2287.76 | 19276.46 ± 3732.43 | 4872.78 ± 849.95 | 41799.69 ± 31552.30 |
6 | 111115.14 ± 32976.86 | 18619.32 ± 5691.99 | 15799.31 ± 5867.25 | 34965.99 ± 11250.10 | 7156.92 ± 1550.86 | 55738.98 ± 40130.05 |
![](/wiki/images/thumb/d/da/K03_summary.jpg/500px-K03_summary.jpg)
Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.
ATP
Time (h) | Mediator | |||||
---|---|---|---|---|---|---|
PGE2 | 15-keto-PGE2 | 13,14-dihydro-15-keto-PGE2 | PGF2α | 12-HETE | 15-HETE | |
0 | 1.50 ± 0.00 | 0.37 ± 0.21 | 2.50 ± 0.00 | 7.50 ± 0.00 | 1.00 ± 0.00 | 2.00 ± 0.00 |
0.5 | 617.77 ± 60.96 | 1.10 ± 0.58 | 51.51 ± 6.47 | 96.88 ± 13.81 | 49.56 ± 37.94 | 15.82 ± 9.10 |
1 | 849.12 ± 56.72 | 3.60 ± 0.79 | 86.88 ± 32.86 | 148.10 ± 20.14 | 32.28 ± 31.72 | 10.95 ± 9.46 |
3 | 833.49 ± 170.07 | 2.05 ± 0.65 | 106.43 ± 12.88 | 170.02 ± 34.73 | 34.05 ± 39.23 | 5.70 ± 7.02 |
6 | 1015.73 ± 88.65 | 2.43 ± 1.70 | 141.16 ± 9.62 | 210.30 ± 43.78 | 9.76 ± 10.52 | 2.22 ± 0.54 |
![](/wiki/images/thumb/c/ca/K04_eicosanoids.jpg/500px-K04_eicosanoids.jpg)
Lipid mediator levels in unstimulated (red) and stimulated with ATP (2mM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.
UVR
Time (h) | Mediator | |||||
---|---|---|---|---|---|---|
PGE2 | 15-keto-PGE2 | 13,14-dihydro-15-keto-PGE2 | PGF2α | 12-HETE | 15-HETE | |
0 | 203.29 ± 315.50 | 0.50 ± 0.00 | 2.00 ± 0.00 | 0.00 ± 0.00 | 3.50 ± 0.00 | 2.30 ± 0.00 |
0.5 | 1.50 ± 0.00 | 0.75 ± 0.39 | 2.00 ± 0.00 | 88.72 ± 33.06 | 135.07 ± 142.32 | 40.09 ± 35.24 |
1 | 69.26 ± 60.99 | 1.05 ± 0.99 | 2.29 ± 1.56 | 90.56 ± 11.96 | 131.60 ± 100.39 | 46.54 ± 35.33 |
3 | 1.50 ± 0.00 | 9.61 ± 7.15 | 3.53 ± 2.41 | 222.44 ± 6.73 | 103.71 ± 78.12 | 58.45 ± 43.57 |
6 | 5667.91 ± 4967.29 | 152.58 ± 51.19 | 6.21 ± 4.30 | 1657.38 ± 297.77 | 766.69 ± 206.41 | 648.45 ± 219.84 |
![](/wiki/images/thumb/4/49/K01_eicosanoids.jpg/500px-K01_eicosanoids.jpg)
Lipid mediator levels in unstimulated (red) and irradiated (15 mJ/cm2) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.
Calcium ionophore + Indomethacin
Time (h) | Mediator | |||||
---|---|---|---|---|---|---|
PGE2 | 15-keto-PGE2 | 13,14-dihydro-15-keto-PGE2 | PGF2α | 12-HETE | 15-HETE | |
0 | 26.81 ± 10.56 | 0.50 ± 0.00 | 2.00 ± 0.00 | 7.00 ± 0.00 | 0.29 ± 0.00 | 1.92 ± 0.94 |
0.5 | 34.23 ± 22.16 | 0.50 ± 0.00 | 1.88 ± 0.57 | 7.00 ± 0.00 | 7.20 ± 10.94 | 29.71 ± 60.25 |
1 | 25.85 ± 9.25 | 0.50 ± 0.00 | 32.91 ± 28.56 | 7.00 ± 0.00 | 141.09 ± 24.66 | 54.23 ± 51.50 |
3 | 4731.85 ± 7300.78 | 2266.13 ± 371.89 | 1817.62 ± 907.18 | 3268.46 ± 658.45 | 17918.03 ± 8595.61 | 55405.98 ± 17910.03 |
6 | 4843.70 ± 7474.03 | 1620.01 ± 317.98 | 900.50 ± 199.18 | 4754.66 ± 596.47 | 61471.62 ± 16768.21 | 105543.44 ± 41314.43 |
![](/wiki/images/thumb/a/a0/K07_eicosanoids.jpg/500px-K07_eicosanoids.jpg)
Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) following 1h incubation with indomethacin (10 mM) (blue) HaCaT keratinocytes, over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.