Table 1.
Targeted lipidome of IL-1β-activated primary HLFs
| Lipid | HLFs + Vehicle | HLFs + IL-1β | P Value (vs. vehicle) | HLFs + IL-1β + Celecoxib | P Value (vs. IL-1β) |
|---|---|---|---|---|---|
| PGE2 | 69.93 ± 20.98 | 3,554.59 ± 264.11 | **** | 4.94 ± 9.22 | **** |
| PGA2 | nd | 332.38 ± 55.58 | *** | nd | *** |
| PGJ2 | 18.05 ± 3.53 | 181.76 ± 25.95 | *** | nd | *** |
| Δ12-PGJ2 | 13.03 ± 3.95 | 598.48 ± 74.84 | *** | 4.82 ± 8.35 | *** |
| PGF2α | nd | 39.58 ± 9.22 | *** | nd | *** |
| 15-keto PGF2α | 2.94 ± 5.10 | 204.92 ± 42.43 | ** | nd | ** |
| TXB2 | nd | 19.27 ± 3.85 | *** | nd | *** |
| 13,14dh-15k-PGD2 | 2.06 ± 3.56 | 19.70 ± 6.93 | * | nd | ** |
| 13,14dh-15k-PGE2 | nd | 174.11 ± 25.17 | *** | nd | *** |
| PGE1 | nd | 41.08 ± 12.41 | ** | nd | *** |
| 15(R)-PGE1 | 7.50 ± 1.54 | 50.40 ± 4.90 | *** | 2.57 ± 4.45 | *** |
| PGF1α | nd | 6.00 ± 0.84 | *** | nd | *** |
| 19(R)-OH PGF2α | 32.80 ± 2.68 | nd | **** | nd | |
| 15d-Δ12,14-PGJ2 | 11.60 ± 3.90 | nd | ** | 2.05 ± 3.54 | |
| 20-COOH LTB4 | 24.77 ± 7.36 | nd | ** | nd | |
| 14,15-diHETrE | 41.36 ± 11.87 | 25.61 ± 1.80 | nd | **** | |
| tetranor 12-HETE | 72.95 ± 9.96 | 34.41 ± 4.94 | ** | 12.38 ± 3.27 | ** |
| 8(S), 15(S)-diHETE | nd | 9.07 ± 15.71 | nd | ||
| 12(13)-diHOME | 34.54 ± 20.58 | 18.44 ± 16.67 | 9.63 ± 13.29 | ||
| 9(10)-epOME | nd | 3.02 ± 12.23 | nd | ||
| 14,15-epETE | 14.60 ± 12.77 | 2.93 ± 5.08 | nd | ||
| 17,18-epETE | 4.81 ± 8.33 | 5.25 ± 9.10 | nd | ||
| 8,9-epETrE | 4.88 ± 8.45 | nd | nd | ||
| 11,12-epETrE | nd | nd | 3.69 ± 6.38 | ||
| 14,15-epETrE | 6.76 ± 11.71 | 5.18 ± 8.97 | 5.48 ± 9.48 | ||
| iPF-VI | 4.66 ± 4.22 | nd | nd | ||
| 12-oxoLTB4 | nd | nd | 6.50 ± 11.25 | ||
| 9(S)-HOTrE | nd | 11.29 ± 59.74 | nd | ||
| 13(S)-HOTrE(g) | nd | 14.63 ± 25.35 | 3.85 ± 6.67 | ||
| 9-HODE | nd | 266.61 ± 934.23 | nd | ||
| 13-HODE | nd | 510.93 ± 1,656.12 | nd | ||
| 15(S)-HEPE | 6.00 ± 6.72 | 1.67 ± 11.01 | 2.39 ± 0.55 | ||
| 5-HETE | nd | 2.28 ± 3.96 | nd | ||
| 8-HETE | 5.40 ± 9.35 | nd | 4.24 ± 7.34 | ||
| 11-HETE | 2.32 ± 4.01 | 9.62 ± 3.17 | 3.40 ± 5.89 | ||
| 8(S)-HETrE | nd | 3.92 ± 6.79 | nd | ||
| 13-HDoHE | 11.90 ± 20.62 | nd | nd | ||
| 20-HDoHE | nd | 2.27 ± 3.94 | nd | ||
| 12-oxoETE | 5.44 ± 5.91 | nd | nd | ||
| 15d-Δ12,14-PGJ3 | nd | 1.65 ± 4.22 | nd | ||
| tetranor PGEM | 1.45 ± 2.51 | nd | 5.70 ± 9.88 | ||
| PGE3 | nd | 5.16 ± 8.94 | nd | ||
| 13,14dh-15k-PGF2α | nd | nd | 48.12 ± 83.35 | ||
| 19(R)-OH PGE1 | 13.62 ± 11.97 | nd | nd | ||
| RvD1 | 10.53 ± 23.10 | nd | 5.94 ± 19.26 |
Values (background-subtracted means ± SD) are presented as lipid concentration in conditioned media (pg/ml); n = 3 independent biological replicates per condition. Primary human lung fibroblasts (HLFs) were treated with IL-1β (0.1 ng/ml) with or without celecoxib (10 μM) for 72 h, and conditioned media were subjected to targeted lipidomics analysis using quantitative LC-MS. Blank medium (i.e., serum-free phenol red-free MEM, 3 replicates) was submitted and analyzed to detect background levels of lipids. Background subtraction was performed on each conditioned medium sample by subtraction of lipid levels of the blanks (mean + 2SD) from each sample measurement. Statistical analysis was performed using actual background-subtracted mediator concentrations; however, when background-subtracted concentrations were zero or negative, the result is listed as not detected (nd):
P < 0.05,
P < 0.01,
P < 0.001,
P < 0.0001 (by unpaired t-test).