Background
Methods
Patients
Patient selection
Combined sample extraction for oxylipins, endocannabinoids and related lipids
LC–MS/MS equipment for analysis of oxylipins, endocannabinoids and related lipids
Standards and calibration curve preparation
Multivariate and univariate data analysis
Results
Characterization of plasma concentration and profiles of oxylipin metabolites
Compound | Abbreviation | Precursor | Pathway | OPLS-DA uncomplicated versus controls | p value uncomplicated versus controls | OPLS-DA severe versus controls | p value severe versus controls | OPLS-DA severe versus uncomplicated | p value severe versus uncomplicated |
---|---|---|---|---|---|---|---|---|---|
Thromboxane B2 | TXB2
| AA | COX | −0.62
| <0.0001
| −0.55
|
0.0004
| 0.13 | NS |
9,12,13-trihydroxy-octadecenoic acid | 9,12,13-TriHOME | LA | 5-LOX | 0.36 | NS | 0.61 | 0.0022 | 0.31 | NS |
9,10,13-trihydroxy-octadecenoic acid | 9,10,13-TriHOME | LA | 5-LOX | 0.27 | NS | 0.61 | 0.0413 | 0.27 | NS |
Prostaglandin F2aα | PGF2α
| AA | COX | −0.10 | NS | 0.00 | 0.0056 | −0.02 | NS |
Prostaglandin E2 | PGE2
| AA | COX | −0.45
| NS | −0.37 | 0.0180 | −0.19 | NS |
Prostaglandin D2 | PGD2
| AA | COX | −0.38
| NS | −0.21 | 0.0428 | −0.03 | NS |
Resolvin D2 | Resolvin D2 | DHA | 12/15-LOX | 0.07 | NS | −0.17 | NS | −0.28 | NS |
Resolvin D1 | Resolvin D1 | DHA | 12/15-LOX | −0.32 | – | −0.16 | – | 0.25 | – |
Lipoxin A4 | 5,6,-LXA4
| AA | 12/15-LOX | −0.19 | NS | −0.10 | NS | −0.00 | NS |
Trans-Leukotriene B4 | Trans-LTB4
| AA | 5-LOX | −0.12 | NS | 0.18 | NS | 0.15 | NS |
Leukotriene B4 | LTB4
| AA | 5-LOX | −0.02 | NS |
0.56
| NS |
0.65
| NS |
12,13-dihydroxy-octadecenoic acid | 12,13-DiHOME | LA | CYP | 0.25 | NS | 0.34 | NS | 0.21 | NS |
9,10-dihydroxy-octadecenoic acid | 9,10-DiHOME | LA | CYP | 0.09 | NS | 0.34 | NS | 0.26 | NS |
14,15-dihydroxy-eicosatrienoic acid | 14,15-DHET | AA | CYP |
0.60
| NS | 0.58 | NS | −0.17 | NS |
11,12-dihydroxy-eicosatrienoic acid | 11,12-DHET | AA | CYP |
0.53
| NS | 0.40 | NS | −0.29 | NS |
8,9-dihydroxy-eicosatrienoic acid | 8,9-DHET | AA | CYP | 0.43 | NS | 0.21 | NS | −0.36 | NS |
5,6-dihydroxy-eicosatrienoic acid | 5,6-DHET | AA | CYP | −0.23 | 0.0054 | −0.26 | 0.0051 | −0.23
| NS |
12-hydroxy-eicosapentaenoic acid | 12-HEPE | EPA | 12/15-LOX | −0.33 | NS | −0.36 | NS | −0.47 | NS |
20-hydroxy-eicosatetraenoic acid | 20-HETE | AA | CYP | 0.37 | 0.0017 | 0.27 | 0.0241 | −0.33 | NS |
13-hydroxy-octadecadienoic acid | 13-HODE | LA | 12/15-LOX | 0.06 | NS | 0.55 | NS | 0.58 | 0.0200 |
9-hydroxy-octadecadienoic acid | 9-HODE | LA | 5-LOX | 0.09 | NS | 0.55 | 0.0350 (*) | 0.57 | 0.0128 (*) |
15-hydroxy-eicosatetraenoic acid | 15-HETE | AA | 12/15-LOX | −0.38 | NS | −0.23 | 0.0402 | −0.25 | NS |
13-oxo-octadecadienoic acid | 13-oxo-ODE | LA | 12/15-LOX | 0.24 | NS | 0.51 | NS |
0.60
| 0.0464 |
11-hydroxy-eicosatetraenoic acid | 11-HETE | AA | 12/15-LOX | −0.37 | NS | −0.26 | NS | −0.19 | NS |
15-oxo-eicosatetraenoic acid | 15-oxo-ETE | AA | 12/15-LOX | −0.08 | NS | 0.29 | NS | 0.32 | NS |
12-hydroxy-eicosatetraenoic acid | 12-HETE | AA | 12/15-LOX | −0.37 | NS | −0.38 | 0.0079 | −0.47 | NS |
8-hydroxy-eicosatetraenoic acid | 8-HETE | AA | 12/15-LOX | −0.13 | NS | 0.02 | NS | −0.11 | NS |
15-hydroxy-eicosatrienoic acid | 15-HETrE | DGLA | 12/15-LOX | −0.27
| 0.0306 | −0.06 | NS | 0.11 | NS |
12-oxo-eicosatetraenoic acid | 12-oxo-ETE | AA | 12/15-LOX | 0.01 | NS | −0.14 | NS | −0.51 | NS |
9-hydroxy-eicosatetraenoic acid | 9-HETE | AA | 12/15-LOX | −0.28 | NS | −0.10 | NS | −0.10 | NS |
5-hydroxy-eicosatetraenoic acid | 5-HETE | AA | 5-LOX | −0.14 | NS | −0.01 | NS | −0.03 | NS |
12(13)epoxy-octadecenoic acid | 12(13)-EpOME | LA | CYP |
0.66
| 0.0087 |
0.72
| 0.0017 | 0.38 | NS |
14(15)-epoxy-eicosatrienoic acid | 14(15)-EET | AA | CYP |
0.60
| NS | 0.60 | NS | 0.26 | NS |
9(10)epoxy-octadecenoic acid | 9(10)-EpOME | LA | CYP |
0.69
| 0.0107 |
0.77
| 0.0025 | 0.39 | NS |
11(12)-epoxy-eicosatrienoic acid | 11(12)-EET | AA | CYP | 0.32 | NS | 0.46 | NS |
0.55
| NS |
5-oxo-eicosatetraenoic acid | 5-oxo-ETE | AA | 5-LOX | 0.09 | NS | 0.27 | NS | 0.01 | NS |
8(9)-epoxy-eicosatrienoic acid | 8(9)-EET | AA | CYP |
0.44
| NS | 0.44 | NS | 0.25 | NS |
5(6)-epoxy-eicosatrienoic acid | 5(6)-EET | AA | CYP | 0.54 | NS | 0.48 | NS | 0.29 | NS |
Univariate analysis of oxylipin levels
ROC curve analysis
Characterization of plasma concentration of endocannabinoids and related endocannabinoid profiles
Compound | Abbreviation | Fatty acid precursor | OPLS-DA uncomplicated versus controls | p value uncomplicated versus controls | OPLS-DA severe versus controls | p value severe versus controls |
---|---|---|---|---|---|---|
2-Arachidonoyl glycerol | 2-AG | AA |
0.70
| 0.0048 |
0.70
|
0.0019
|
Arachidonoyl ethanolamide | AEA | AA | 0.20 | NS | 0.43 | NS |
Oleoyl ethanolamide | OEA | OA |
0.88
| <0.0001
|
0.97
| <0.0001
|
Palmitoyl ethanolamide | PEA | PA |
0.80
|
0.0018
|
0.86
|
0.0001
|
Docosatetraenoyl ethanolamide | DEA | DTA |
0.82
|
0.0009
|
0.83
|
0.0002
|
Arachidonoyl glycine | NAGLy | AA | −0.52 | NS |
−0.40
| NS |
Eicosapentaenoyl ethanolamide | EPEA | EPA |
0.57
|
0.0017(*) |
0.56
|
0.0006
|
Docosahexaenoyl ethanolamide | DHEA | DHA |
0.56
| 0.0046 |
0.69
|
0.0025
|
Palmitoleoyl ethanolamide | POEA | PO | 0.57 | 0.0145 |
0.52
| 0.0224 |
Linolenoyl ethanolamide | LEA | LA | 0.36 | NS | 0.26 | NS |
Prostaglandin F2α ethanolamide | PGF2α EA | AA | −0.28 | NS | 0.02 | NS |
Prostaglandin E2 ethanolamide | PGE2 EA | AA | 0.03 | NS | 0.15 | NS |