Table 3 SPM profiles in IL-1β stimulated diseased tendon stromal cells incubated in 15-epi-LXA4 and inhibitors of 15-PGDH.

From: Increased 15-PGDH expression leads to dysregulated resolution responses in stromal cells from patients with chronic tendinopathy

DHA bioactive metabolome

Q1

Q3

Tendon stromal cells Lipid mediators levels (pg/incubation)

Disease + IL-1β

Disease + IL-1β + 15-epi-LXA4

Disease + IL-1β + Indo

Disease + IL-1β + 15-epi-LXA4 + Indo

Disease + IL-1β + SW033291

Disease + IL-1β + 15-epi-LXA4 + SW033291

RvD1

375

141

0.2 ± 0.2

1.0 ± 1.0

—

—

—

—

RvD2

375

141

0.6 ± 0.4

1.3 ± 0.3

—

—

0.1 ± 0.1

—

RvD3

375

147

0.3 ± 0.3

0.2 ± 0.1

—

—

—

—

RvD4

375

101

1.3 ± 1.3

1.9 ± 0.2

—

—

—

—

RvD5

359

199

34.0 ± 3.8

27.5 ± 5.2

1.3 ± 1.3**

0.3 ± 0.3**

8.6 ± 1.2**

2.1 ± 1.2**

RvD6

359

101

14.9 ± 2.5

20.4 ± 0.4*

4.1 ± 0.5**

4.1 ± 0.2**

3.1 ± 1.3**

1.0 ± 0.7**

17R-RvD1

375

141

1.2 ± 0.7

2.4 ± 1.3

—

0.3 ± 0.3

—

—

17R-RvD3

375

147

2.5 ± 1.5

4.5 ± 1.1

— *

—

—

—

PD1

359

153

7.0 ± 1.1

7.0 ± 1.0

1.5 ± 1.0*

0.8 ± 0.7**

1.4 ± 0.7**

0.7 ± 0.7**

17R-PD1

359

153

0.2 ± 0.1

0.3 ± 0.3

—

0.0 ± 0.0

0.1 ± 0.1

0.3 ± 0.2

10 S,17S-diHDHA

359

153

34.6 ± 5.2

64.3 ± 6.2*

0.6 ± 0.6*

1.8 ± 1.8**

0.9 ± 0.9**

—**

MaR1

359

221

0.9 ± 0.9

13.2 ± 2.1**

0.0 ± 0.0

1.2 ± 1.2

0.7 ± 0.7

—

MaR2

359

191

0.5 ± 0.3

1.0 ± 0.1

0.7 ± 0.3

1.0 ± 0.6

2.1 ± 0.5*

—

7 S,14S-diHDHA

359

221

32.0 ± 5.2

22.1 ± 6.7

—

0.4 ± 0.4**

—**

—**

4 S,14S-diHDHA

359

101

—

—

—

—

—

—

14-oxo-MaR1

357

248

0.3 ± 0.2

0.4 ± 0.4

0.3 ± 0.2

0.1 ± 0.1

0.2 ± 0.2

—

n-3 DPA bioactive metabolome

RvD1n-3 DPA

377

143

3.2 ± 1.9

4.7 ± 0.6

0.1 ± 0.1

0.2 ± 0.2

—

—

RvD2n-3 DPA

377

261

0.4 ± 0.3

—

—

0.0 ± 0.0

—

—

RvD5n-3 DPA

361

263

25.4 ± 1.7

14.6 ± 2.4 *

— **

1.2 ± 1.2

— **

— **

PD1n-3 DPA

361

183

0.9 ± 0.5

2.2 ± 1.1

0.4 ± 0.4

0.8 ± 0.2

4.2 ± 1.8

0.9 ± 0.3

10 S,17S-diHDPA

361

183

16.8 ± 1.7

16.8 ± 2.3

5.2 ± 1.3 *

6.0 ± 0.7 **

3.3 ± 0.5 **

3.3 ± 1.0 **

MaR1n-3 DPA

361

249

3.9 ± 2.1

2.8 ± 0.6

0.4 ± 0.4

0.4 ± 0.2

0.1 ± 0.1

—

EPA bioactive metabolome

RvE1

349

195

—

—

—

0.0 ± 0.0

— **

0.1 ± 0.1

RvE2

333

199

49.0 ± 11.7

44.3 ± 9.4

18.9 ± 2.0 *

22.3 ± 4.6 *

23.6 ± 4.0 *

24.6 ± 4.0

RvE3

333

201

0.4 ± 0.3

0.0 ± 0.0

—

0.0 ± 0.0

—

—

AA bioactive metabolome

LXA4

351

217

—

15.9 ± 10.6

—

4.9 ± 0.5** $

—

2.4 ± 1.4

LXB4

351

221

1.4 ± 1.4

—

—

0.0 ± 0.0

—

—

5 S,15S-diHETE

335

235

121.8 ± 14.8

770.0 ± 168.4**

— *

2.9 ± 2.9*

— **

—

15epi-LXA4

351

217

—

253.2 ± 16.3**

—

291 ± 20.2**$$

—

297.1 ± 23.9**##

15epi-LXB4

351

221

35.0 ± 7.3

45.5 ± 6.6

18.5 ± 3.6*

21.3 ± 5.9

— **

5.4 ± 2.7*

15-oxo-LXA4

349

233

—

3.0 ± 0.5*

—

0.6 ± 0.1$$

0.1 ± 0.0

0.5 ± 0.2* #

LTB4

335

195

—

 

—

   

5 S,12S-diHETE

335

195

45.6 ± 45.6

40.9 ± 31.1

—

0.0 ± 0.0

—

—

PGD2

351

189

50.0 ± 4.0

61.0 ± 9.4

0.7 ± 0.7**

30.8 ± 3.8* #

1.2 ± 1.2 **

29.4 ± 6.0*#

PGE2

351

189

1655.4 ± 108.7

1275.0 ± 95.0 **

4.6 ± 3.6**

4.8 ± 3.8**

154.5 ± 14.8**

74.6 ± 20.7** #

PGF2α

353

193

258.1 ± 26.0

258.8 ± 51.3

—**

0.0 ± 0.0**

2.4 ± 2.2**

5.7 ± 5.3**

TxB2

369

169

374.8 ± 31.5

367.9 ± 77.2

— **

0.0 ± 0.0**

3.3 ± 3.0**

7.9 ± 7.4**

  1. Tendon stromal cells (60,000 cells per well) were derived from diseased supraspinatus tendons. Cells were incubated for 24 h (37 °C) with indomethacin (indo; 10 µM), SW033291 (25 µM) or vehicle (2 h, 37 °C), then with 15-epi-LXA4 (10 nM) or vehicle (24 h 37 °C) and with IL-1β (37 °C; 24 h). 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 = 3 per incubation. *P < 0.05, **P < 0.01 vs Disease + IL1β incubations. $P < 0.05, $$P < 0.01 vs Disease + IL1β  + Indo; #P < 0.05, ##P < 0.01 vs Disease + IL1β  + SW. The detection limit was ~ 0.1 pg. -, Below levels found in media alone.
Back to article page