Table 1.
Altered LM profiles of diseased tendon stromal cells when compared to those from healthy tendon stromal cells.
| DHA bioactive metabolome | Q1 | Q3 | Tendon stromal cells Lipid mediators levels (pg/incubation) | |
|---|---|---|---|---|
| Healthy + IL-1β | Disease + IL-1β | |||
| RvD1 | 375 | 141 | 5.3 ± 1.8 | 4.6 ± 2.0 |
| RvD2 | 375 | 141 | 8.7 ± 4.3 | 11.6 ± 5.3 |
| RvD3 | 375 | 147 | 4.1 ± 2.5 | 6.6 ± 2.7 |
| RvD4 | 375 | 101 | 4.2 ± 2.1 | 7.9 ± 3.3 |
| RvD5 | 359 | 199 | 53.9 ± 11.1 | 49.3 ± 7.1 |
| RvD6 | 359 | 101 | 24.1 ± 7.7 | 13.9 ± 2.7 |
| 17R-RvD1 | 375 | 141 | 1.9 ± 0.6 | 2.1 ± 0.6 |
| 17R-RvD3 | 375 | 147 | 5.7 ± 1.7 | 7.6 ± 2.1 |
| PD1 | 359 | 153 | 27.6 ± 5.8 | 26.6 ± 9.5 |
| 17R-PD1 | 359 | 153 | 1.0 ± 0.2 | 1.7 ± 1.0 |
| 10 S,17S-diHDHA | 359 | 153 | 139.7 ± 10.3 | 117.1 ± 41.3 |
| MaR1 | 359 | 221 | 6.3 ± 2.4 | 28.7 ± 11.8* |
| 7 S,14S-diHDHA | 359 | 221 | 40.5 ± 22.3 | 7.9 ± 2.9 |
| 4 S,14S-diHDHA | 359 | 101 | 15.8 ± 6.8 | 19.8 ± 6.9 |
| n-3 DPA bioactive metabolome | ||||
| RvD1n-3 DPA | 377 | 143 | 2.0 ± 0.8 | 4.8 ± 1.0* |
| RvD2n-3 DPA | 377 | 261 | 0.7 ± 0.6 | 2.3 ± 1.3 |
| RvD5n-3 DPA | 361 | 263 | 19.9 ± 4.3 | 23.0 ± 5.9 |
| PD1n-3 DPA | 361 | 183 | 58.0 ± 26.4 | 98.6 ± 54.0 |
| 10 S,17S-diHDPA | 361 | 183 | 414.3 ± 207.1 | 499.3 ± 263.5 |
| MaR1n-3 DPA | 361 | 249 | 13.8 ± 5.6 | 13.3 ± 4.5 |
| EPA bioactive metabolome | ||||
| RvE1 | 349 | 195 | 0.3 ± 0.2 | 0.1 ± 0.1 |
| RvE2 | 333 | 199 | 39.8 ± 10.1 | 54.3 ± 6.0 |
| RvE3 | 333 | 201 | 2.0 ± 1.1 | 2.0 ± 0.9 |
| AA bioactive metabolome | ||||
| LXA4 | 351 | 217 | 2.2 ± 1.0 | 6.0 ± 1.7* |
| LXB4 | 351 | 221 | 40.9 ± 18.5 | 96.2 ± 52.9 |
| 5 S,15S-diHETE | 335 | 235 | 1404.0 ± 358.7 | 1727.2 ± 437.5 |
| 15epi-LXA4 | 351 | 217 | 82.8 ± 48.4 | 245.2 ± 78.3* |
| 15epi-LXB4 | 351 | 221 | 57.7 ± 16.8 | 43.6 ± 12.4 |
| 15-oxo-LXA4 | 349 | 233 | 0.5 ± 0.2 | 3.0 ± 1.3* |
| LTB4 | 335 | 195 | 57.0 ± 49.4 | 0.0 ± 0.0 |
| 5 S,12S-diHETE | 335 | 195 | 138.8 ± 63.9 | 264.8 ± 88.0 |
| PGD2 | 351 | 189 | 2065.8 ± 906.4 | 968.0 ± 294.1 |
| PGE2 | 351 | 189 | 9483.0 ± 4429.1 | 21371.2 ± 8192.9 |
| PGF2α | 353 | 193 | 367.0 ± 79.8 | 380.1 ± 102.5 |
| TxB2 | 369 | 169 | 20.8 ± 5.8 | 12.1 ± 2.2 |
Tendon stromal cells (60,000 cells per well) were derived from healthy hamstring (n = 8 donors) or diseased supraspinatus tendons (n = 8 donors). Cells were incubated for 24 h with IL-1β, incubations were quenched using ice-cold methanol containing deuterium labelled internal standards and lipid mediators (LM) were identified and quantified using LM profiling (see methods for details). Q1, M-H (parent ion) and Q3, diagnostic ion in the MS-MS (daughter ion). Results are expressed as pg/incubation. Mean ± SEM of n = 8 per incubation. *P < 0.05 vs Healthy. The detection limit was ~ 0.1 pg. -, Below levels found in media alone.