#METABOLOMICS WORKBENCH michaelsa93_20170809_103526 DATATRACK_ID:1194 STUDY_ID:ST000843 ANALYSIS_ID:AN001365 PROJECT_ID:PR000600 VERSION 1 CREATED_ON August 10, 2017, 2:42 pm #PROJECT PR:PROJECT_TITLE Statin Immuno-Metabolomics in Asthma PR:PROJECT_TYPE Placebo-controled trial PR:PROJECT_SUMMARY Innovative and novel therapies are urgently needed for the treatment of patients PR:PROJECT_SUMMARY with severe asthma, especially those who are refractory to standard-of-care PR:PROJECT_SUMMARY bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the PR:PROJECT_SUMMARY role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation PR:PROJECT_SUMMARY and remodeling. Although statins all inhibit HMGCR in the same manner in terms PR:PROJECT_SUMMARY of enzyme binding site, the statins’ varied physiochemical properties with PR:PROJECT_SUMMARY respect to their polarity (i.e. lipophilicity) result in very different immune PR:PROJECT_SUMMARY and lipid effects. The major significance of this work is to advance a new class PR:PROJECT_SUMMARY of inhaler therapies for asthma; the statins which work by an entirely different PR:PROJECT_SUMMARY mechanism than current ICS/LABA mainstays. Evidence suggests that statins may PR:PROJECT_SUMMARY have an additive benefit to corticosteroids in asthma, thereby confirming a PR:PROJECT_SUMMARY unique mechanism, namely via MVA pathway inhibition. This becomes particularly PR:PROJECT_SUMMARY important in the severe asthma population which is highly PR:PROJECT_SUMMARY corticosteroid-resistant, is poorly controlled with high exacerbation rates and PR:PROJECT_SUMMARY hospitalizations, and has the highest healthcare costs of all asthma phenotypes. PR:PROJECT_SUMMARY In essence, the potential public health impact of even an incremental PR:PROJECT_SUMMARY improvement in asthma symptom control cannot be underestimated. Even the PR:PROJECT_SUMMARY prevention of 1 asthma attack preserves lung function and reduces the adverse PR:PROJECT_SUMMARY personal and financial impact. This study aimed to determine if statin polarity PR:PROJECT_SUMMARY affects airway drug concentration and systemic drug absorption and to determine PR:PROJECT_SUMMARY the effect of inhaled statins on naïve airway immune cell populations and PR:PROJECT_SUMMARY alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus PR:PROJECT_SUMMARY monkeys. In this particular component of the study, we investigated the PR:PROJECT_SUMMARY metabolic effects resulting from the use of statins in these healthy rhesus PR:PROJECT_SUMMARY monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, PR:PROJECT_SUMMARY endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, PR:PROJECT_SUMMARY plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma. PR:INSTITUTE University of California, Davis PR:DEPARTMENT Internal Medicine PR:LAST_NAME Zeki PR:FIRST_NAME Amir PR:ADDRESS 2825 J St. Suite 400 Sacramento, CA 95816 PR:EMAIL aazeki@ucdavis.edu PR:PHONE (916) 734-8230 #STUDY ST:STUDY_TITLE Statin Immuno-Metabolomics in Asthma (part I) ST:STUDY_TYPE Placebo-controled trial ST:STUDY_SUMMARY Innovative and novel therapies are urgently needed for the treatment of patients ST:STUDY_SUMMARY with severe asthma, especially those who are refractory to standard-of-care ST:STUDY_SUMMARY bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the ST:STUDY_SUMMARY role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation ST:STUDY_SUMMARY and remodeling. Although statins all inhibit HMGCR in the same manner in terms ST:STUDY_SUMMARY of enzyme binding site, the statins’ varied physiochemical properties with ST:STUDY_SUMMARY respect to their polarity (i.e. lipophilicity) result in very different immune ST:STUDY_SUMMARY and lipid effects. The major significance of this work is to advance a new class ST:STUDY_SUMMARY of inhaler therapies for asthma; the statins which work by an entirely different ST:STUDY_SUMMARY mechanism than current ICS/LABA mainstays. Evidence suggests that statins may ST:STUDY_SUMMARY have an additive benefit to corticosteroids in asthma, thereby confirming a ST:STUDY_SUMMARY unique mechanism, namely via MVA pathway inhibition. This becomes particularly ST:STUDY_SUMMARY important in the severe asthma population which is highly ST:STUDY_SUMMARY corticosteroid-resistant, is poorly controlled with high exacerbation rates and ST:STUDY_SUMMARY hospitalizations, and has the highest healthcare costs of all asthma phenotypes. ST:STUDY_SUMMARY In essence, the potential public health impact of even an incremental ST:STUDY_SUMMARY improvement in asthma symptom control cannot be underestimated. Even the ST:STUDY_SUMMARY prevention of 1 asthma attack preserves lung function and reduces the adverse ST:STUDY_SUMMARY personal and financial impact. This study aimed to determine if statin polarity ST:STUDY_SUMMARY affects airway drug concentration and systemic drug absorption and to determine ST:STUDY_SUMMARY the effect of inhaled statins on naïve airway immune cell populations and ST:STUDY_SUMMARY alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus ST:STUDY_SUMMARY monkeys. In this particular component of the study, we investigated the ST:STUDY_SUMMARY metabolic effects resulting from the use of statins in these healthy rhesus ST:STUDY_SUMMARY monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, ST:STUDY_SUMMARY endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, ST:STUDY_SUMMARY plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma. ST:INSTITUTE USDA ST:DEPARTMENT Obesity and metabolism research unit ST:LABORATORY Newman's Lab ST:LAST_NAME Newman ST:FIRST_NAME John ST:ADDRESS 430 West Health Sciences Dr. Davis, Ca, 95616 ST:EMAIL John.Newman@ars.usda.gov ST:PHONE (530) 752-1009 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Macaca mulatta SU:TAXONOMY_ID 9544 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - BAL-AZ-01 Treatment:Control | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-33 Treatment:Control | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-06 Treatment:Simvastatin | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-24 Treatment:Simvastatin | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-11 Treatment:Simvastatin | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-10 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-09 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-04 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-28 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-08 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-23 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-19 Treatment:Control | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-34 Treatment:Control | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-02 Treatment:Control | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-05 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-35 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-25 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-18 Treatment:Control | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-07 Treatment:Control | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-30 Treatment:Simvastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-27 Treatment:Simvastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-03 Treatment:Simvastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - BAL-AZ-14 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-17 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-13 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-32 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-15 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-29 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - BAL-AZ-26 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-12 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-22 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-20 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-31 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - BAL-AZ-21 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 01 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 02 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 03 Treatment:Control | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 06 Treatment:Control | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 07 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 08 Treatment:Pravastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 09 Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 10 Treatment:Control | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 13 Treatment:Control | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 14 Treatment:Pravastatin | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 15 Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 16 Treatment:Control | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 17 Treatment:Pravastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 18 Treatment:Pravastatin | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 21 Treatment:Pravastatin | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 22 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 23 Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 24 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 25 Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 26 Treatment:Pravastatin | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 29 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 30 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 31 Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 32 Treatment:Control | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 33 Treatment:Control | Lung:Left Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 36 Treatment:Control | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 37 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 38 Treatment:Control | Lung:Right Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 39 Treatment:Control | Lung:Right Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 40 Treatment:Pravastatin | Lung:Right Lung | Time point (day):- SUBJECT_SAMPLE_FACTORS - AZ 43 Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 SUBJECT_SAMPLE_FACTORS - AZ 44 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 SUBJECT_SAMPLE_FACTORS - AZ 45 Treatment:Control | Lung:Left Lung | Time point (day):12 #COLLECTION CO:COLLECTION_SUMMARY Monkeys were treated with placebo or Provastatin for 12 days. Further, after the CO:COLLECTION_SUMMARY wash out period animals were treated with Simvastatin for 12 days. Lung wash CO:COLLECTION_SUMMARY fluid was collected at day 0, 8 and 12 of each treatment. CO:SAMPLE_TYPE Bronchoalveolar lavage #TREATMENT TR:TREATMENT_SUMMARY Monkeys were treated (by inhalation) with placebo or Provastatin for 12 days. TR:TREATMENT_SUMMARY Further, after the wash out period animals were treated with Simvastatin for 12 TR:TREATMENT_SUMMARY days. Lung wash fluid was collected at day 0, 8 and 12 of each treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Oxylipins, endocannabinoids, and fatty acids were isolated by solid phase SP:SAMPLEPREP_SUMMARY extraction on 60 mg Waters Oasis-HLB cartridges (Milford, MA), as previously SP:SAMPLEPREP_SUMMARY described by Luria et al (1). Prior to extraction, cartridges were washed with 1 SP:SAMPLEPREP_SUMMARY column volume ethyl acetate followed by 2 column volumes methanol and SP:SAMPLEPREP_SUMMARY conditioned with 2 mL of 95:5 v/v water/methanol (MeOH) with 0.1% acetic acid. SP:SAMPLEPREP_SUMMARY The column reservoir was spiked with 5 µL anti-oxidant solution, (0.2 mg/ml SP:SAMPLEPREP_SUMMARY solution BHT/EDTA in 1:1 MeOH:water), and 10 μL 1000nM analytical surrogates SP:SAMPLEPREP_SUMMARY (See Table 2 below for specific compounds). Sample aliquots (500 µL media) were SP:SAMPLEPREP_SUMMARY then introduced to the column reservoir and diluted with 1 column volume wash SP:SAMPLEPREP_SUMMARY solution (5% MeOH, 0.1% acetic acid) and allowed to gravity extract into tubes SP:SAMPLEPREP_SUMMARY containing 10% bleach solution to decontaminate sample waste. SPE cartridges SP:SAMPLEPREP_SUMMARY were dried by vacuum @ -7.5in Hg for 20 min. Analytes were then eluted by SP:SAMPLEPREP_SUMMARY gravity with 0.5 mL MeOH, followed by 1.25 mL Ethyl Acetate, into 2 mL SP:SAMPLEPREP_SUMMARY autosampler vials containing 10 µL 20% glycerol solution in MeOH. Eluent was SP:SAMPLEPREP_SUMMARY dried by vacuum evaporation for 35 min, and residues were re-constituted with SP:SAMPLEPREP_SUMMARY 100uL of 100 nM internal standard solution containing 1-cyclohexyl SP:SAMPLEPREP_SUMMARY ureido,3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU), in SP:SAMPLEPREP_SUMMARY 50:50 MeOH:ACN and let sit for 10 min at ambient temperature. Vials were then SP:SAMPLEPREP_SUMMARY chilled 15 min on wet ice, and extracts were transferred to a centrifugal filter SP:SAMPLEPREP_SUMMARY (0.1 µm Durapore, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at SP:SAMPLEPREP_SUMMARY <4500g (rcf) and transferred to 150 uL glass inserts within 2 mL amber vials, SP:SAMPLEPREP_SUMMARY and capped. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The SP:SAMPLEPREP_SUMMARY internal standard was used to quantify the recovery of the deuterated extraction SP:SAMPLEPREP_SUMMARY surrogates by ratio response. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (100 x 2mm, 1.7um) CH:FLOW_GRADIENT See protocol/methods file CH:FLOW_RATE 0.25 mL/min CH:COLUMN_TEMPERATURE 60 °C CH:SOLVENT_A 0.1% acetic acid CH:SOLVENT_B 90% ACN / 10% IPA CH:INTERNAL_STANDARD See protocol/methods file CH:RETENTION_TIME See protocol/methods file CH:SAMPLE_INJECTION 5 µL CH:ANALYTICAL_TIME 20 min CH:WEAK_WASH_SOLVENT_NAME 20% methanol, 10% isopropanol CH:WEAK_WASH_VOLUME 600 µL CH:STRONG_WASH_SOLVENT_NAME 50:50 Acetonitrile:Methanol CH:STRONG_WASH_VOLUME 600 µL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME ABI Sciex 6500 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Concentration (nM) MS_METABOLITE_DATA_START Samples BAL-AZ-01 BAL-AZ-33 BAL-AZ-06 BAL-AZ-24 BAL-AZ-11 BAL-AZ-10 BAL-AZ-09 BAL-AZ-04 BAL-AZ-28 BAL-AZ-08 BAL-AZ-23 BAL-AZ-19 BAL-AZ-34 BAL-AZ-02 BAL-AZ-05 BAL-AZ-35 BAL-AZ-25 BAL-AZ-18 BAL-AZ-07 BAL-AZ-30 BAL-AZ-27 BAL-AZ-03 BAL-AZ-14 BAL-AZ-17 BAL-AZ-13 BAL-AZ-32 BAL-AZ-15 BAL-AZ-29 BAL-AZ-26 BAL-AZ-12 BAL-AZ-22 BAL-AZ-20 BAL-AZ-31 BAL-AZ-21 AZ 01 AZ 02 AZ 03 AZ 06 AZ 07 AZ 08 AZ 09 AZ 10 AZ 13 AZ 14 AZ 15 AZ 16 AZ 17 AZ 18 AZ 21 AZ 22 AZ 23 AZ 24 AZ 25 AZ 26 AZ 29 AZ 30 AZ 31 AZ 32 AZ 33 AZ 36 AZ 37 AZ 38 AZ 39 AZ 40 AZ 43 AZ 44 AZ 45 Factors Treatment:Control | Lung:Left Lung | Time point (day):- Treatment:Control | Lung:Left Lung | Time point (day):- Treatment:Simvastatin | Lung:Left Lung | Time point (day):- Treatment:Simvastatin | Lung:Left Lung | Time point (day):- Treatment:Simvastatin | Lung:Left Lung | Time point (day):- Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 Treatment:Simvastatin | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):12 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 Treatment:Simvastatin | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):- Treatment:Control | Lung:Right Lung | Time point (day):- Treatment:Simvastatin | Lung:Right Lung | Time point (day):- Treatment:Simvastatin | Lung:Right Lung | Time point (day):- Treatment:Simvastatin | Lung:Right Lung | Time point (day):- Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 Treatment:Simvastatin | Lung:Right Lung | Time point (day):8 Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 Treatment:Simvastatin | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Right Lung | Time point (day):- Treatment:Control | Lung:Right Lung | Time point (day):- Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Pravastatin | Lung:Right Lung | Time point (day):- Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):12 Treatment:Pravastatin | Lung:Right Lung | Time point (day):8 Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):- Treatment:Pravastatin | Lung:Right Lung | Time point (day):- Treatment:Pravastatin | Lung:Right Lung | Time point (day):8 Treatment:Pravastatin | Lung:Left Lung | Time point (day):- Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 Treatment:Pravastatin | Lung:Left Lung | Time point (day):- Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Pravastatin | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):- Treatment:Control | Lung:Left Lung | Time point (day):- Treatment:Control | Lung:Left Lung | Time point (day):8 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Right Lung | Time point (day):12 Treatment:Control | Lung:Right Lung | Time point (day):8 Treatment:Pravastatin | Lung:Right Lung | Time point (day):- Treatment:Pravastatin | Lung:Left Lung | Time point (day):12 Treatment:Pravastatin | Lung:Left Lung | Time point (day):8 Treatment:Control | Lung:Left Lung | Time point (day):12 1-OG 0.613 0.4 0.29 1.47 0.228 1.11 0.381 0.168 0.711 0.409 0.881 1.31 0.817 0.454 0.355 0.335 0.785 0.273 0.251 0.662 0.702 0.28 0.286 0.243 0.258 0.818 0.25 0.631 0.838 0.413 0.571 1.02 0.642 1.2 0.314 1.19 1.67 0.995 0.78 0.878 0.778 0.916 0.93 0.451 0.914 0.259 0.962 0.698 1.24 0.999 0.586 0.671 0.95 1.6 3.41 2.06 0.599 0.59 0.683 1.01 0.438 0.146 0.948 3.12 0.817 0.48 0.329 2-OG 4.1 0.739 1.8 2.29 0.741 6.67 3.7 0.824 2.87 0.526 1.45 3.29 2.18 1.67 1.62 0.691 2.28 0.723 1.64 1.35 0.859 1.07 1.1 0.737 0.545 5.55 0.533 2.09 2.84 1.71 0.501 1.95 0.606 1.14 0.942 3.23 3.56 6.12 1.48 2.77 1.63 2.91 2.73 1.01 1.37 0.678 1.46 2.35 6.77 1.8 0.756 1.52 2.87 2.87 6.91 6.14 1.6 1.29 1.34 3.55 1.3 1.34 2.46 5.54 1.83 3.52 0.816 1-LG 0.314 0.249 0.291 1.07 0.321 0.841 0.112 0.14 1.14 0.168 0.194 0.333 0.376 0.247 0.249 0.123 0.537 0.15 0.128 0.279 0.185 0.335 0.117 0.112 0.0813 0.397 0.21 0.599 0.464 0.419 0.334 0.837 0.165 0.524 0.193 1.44 0.958 1.44 0.332 0.291 0.474 0.264 0.864 0.296 0.713 1.37 1.24 0.649 1.07 0.451 0.43 0.328 1.19 0.738 1.92 1.04 0.863 0.452 0.227 0.552 0.3 1.91 0.51 1.61 0.43 1.73 0.84 2-LG 3.5 1.11 3.97 3.21 1.73 9.64 5.86 1.57 5.3 1.03 1.55 6.15 2.43 2.55 3.42 1.05 3.72 1.71 2.4 2.6 1.62 1.75 1.84 0.627 0.87 8 1.26 2.27 4.07 3.78 0.975 3.33 1.13 1.96 1.34 3.79 2.66 4.9 1.96 2.85 2.38 3.01 3.26 1.74 2.03 2.25 2.43 3.53 7.1 2.19 1.45 1.86 4.07 3.2 6.42 7.46 3.56 2.42 1.85 2.2 2.26 19.1 1.03 3.85 2.34 4.14 3.12 1-AG 0.0675 0.0502 0.0365 0.145 0.083 0.12 0.0295 0.0195 0.129 0.0393 0.0353 0.0879 0.0707 0.0311 0.0657 0.0149 0.08 0.0389 0.0167 0.0298 0.0264 0.0756 0.0187 0.0177 0.016 0.0586 0.0289 0.0723 0.0423 0.105 0.0553 0.0868 0.0269 0.0614 0.0336 0.576 0.362 0.284 0.0459 0.0859 0.115 0.0628 0.236 0.131 0.23 0.4 0.232 0.234 0.291 0.164 0.0968 0.0536 0.365 0.192 0.187 0.355 0.414 0.0758 0.089 0.119 0.111 0.144 0.116 0.409 0.14 0.176 0.151 2-AG 1.42 0.349 1.57 1.19 0.661 2.35 1.34 0.442 1.54 0.249 0.47 1.46 0.927 0.78 1.38 0.319 1.09 1.1 0.695 0.627 0.408 1.12 0.332 0.483 0.521 2.18 0.387 0.632 0.386 1.04 0.388 0.995 0.383 0.702 0.492 2.01 1.1 1.62 0.528 0.748 1.16 1.03 0.991 0.816 0.777 1.27 0.479 1.44 3.09 1.74 1.05 0.697 1.63 1.15 1.05 2.65 2.28 0.736 0.573 0.961 0.989 0.93 0.515 1.77 1.83 0.64 0.91 PEA 3.66 3.61 6.34 7.81 5 4.33 9.44 9.26 1.35 7.61 7.3 6.09 3.33 3.6 8.12 3.95 1.28 3.27 8.89 5.94 4.1 9.5 7.97 7.04 4.77 8.72 11.5 6.24 2.66 9.87 9.1 8.99 6.92 11.2 0.608 0.631 1.88 0.628 1.45 0.493 0.537 0.546 0.582 0.792 0.802 0.757 0.662 0.478 0.673 0.724 0.693 1.48 0.638 0.759 0.507 0.501 0.77 0.599 1.25 0.711 0.869 0.45 0.452 0.878 0.662 0.632 0.798 SEA 7.42 1.4 10.3 9.9 4.21 4.7 6.37 4.43 1.03 3.45 3.86 10.5 3.92 7.54 5.22 4.96 5.21 4.56 5.9 0.579 4.68 8.76 9.59 3.06 8.07 14.9 7.35 1.16 3.65 5.04 15.6 2.46 7.34 0.29 0.413 1.77 0.132 0.61 0.458 0.535 0.457 0.613 0.378 1.08 0.161 0.464 0.282 0.391 0.882 0.218 1.42 0.624 0.394 0.658 0.589 0.441 0.372 0.569 0.421 0.194 0.08 0.399 0.214 0.76 0.149 0.149 OEA 0.0671 0.0156 0.0497 0.0734 0.0235 0.0967 0.0317 0.0238 0.054 0.0442 0.0593 0.059 0.0298 0.04 0.0621 0.0295 0.0499 0.0286 0.0204 0.037 0.0251 0.0358 0.026 0.0298 0.0119 0.0486 0.0323 0.0374 0.0233 0.0265 0.0288 0.0631 0.0274 0.044 0.0085 0.0326 0.0419 0.0819 0.0318 0.0397 0.0462 0.0325 0.0252 0.0277 0.0424 0.048 0.0337 0.0421 0.0599 0.0185 0.0307 0.0362 0.0416 0.0671 0.0271 0.0519 0.0249 0.0415 0.0205 0.0369 0.0402 0.0353 0.0243 0.0609 0.0386 0.0414 0.0306 LEA 0.0147 0.0039 0.011 0.0219 0.0082 0.0231 0.009 0.0036 0.0145 0.0095 0.013 0.0155 0.0099 0.0077 0.0144 0.0068 0.02 0.0101 0.0048 0.0091 0.0092 0.0087 0.006 0.0049 0.0018 0.0119 0.0066 0.0108 0.0068 0.0064 0.0041 0.0138 0.0073 0.0136 0.0034 0.0156 0.0126 0.0395 0.0121 0.0161 0.0191 0.0152 0.0116 0.0121 0.0209 0.0222 0.0147 0.0141 0.0237 0.0105 0.0155 0.0112 0.0178 0.0292 0.0123 0.0224 0.0099 0.0164 0.0134 0.0196 0.0172 0.0149 0.0151 0.0473 0.0117 0.0185 0.0113 aLEA 0.0003 0.0002 0.0004 0.0008 0.0003 0.0004 0.0002 0.0002 0.0004 0.0002 0.0003 0.0004 0.0003 0.0002 0.0004 0.0002 0.0005 0.0002 0.0001 0.0003 0.0004 0.0004 0.0001 0.0001 0.0001 0.0002 0.0002 0.0003 0.0001 0.0002 0.0002 0.0004 0.0002 0.0004 Dihomo GLA EA 0.0046 0.0005 0.0017 0.0032 0.0006 0.0058 0.0011 0.0002 0.0013 0.0009 0.0017 0.0034 0.002 0.0009 0.0014 0.0005 0.0023 0.0014 0.0006 0.0013 0.0011 0.0009 0.0009 0.0011 0.0001 0.0012 0.0005 0.0007 0.0016 0.0008 0.0005 0.0015 0.0006 0.0013 0.0007 0.001 0.001 0.0015 0.0016 0.0015 0.0012 0.0055 0.0021 0.0006 0.0024 0.0047 0.0007 0.0002 0.0036 0.0019 0.0006 0.0002 0.0022 0.004 0.0012 0.0031 0.0011 0.0007 0.0003 0.0028 0.0018 0.0021 0.0021 0.0002 0.0013 0.0011 0.0009 AEA 0.0115 0.0012 0.003 0.0083 0.0024 0.0107 0.004 0.0003 0.0032 0.0035 0.0042 0.0089 0.0049 0.0019 0.0035 0.0022 0.008 0.0043 0.0015 0.0032 0.0028 0.0024 0.0018 0.0026 0.0002 0.0024 0.0021 0.0025 0.0031 0.0024 0.0009 0.0033 0.0026 0.0039 0.0006 0.0057 0.0049 0.0241 0.0017 0.0061 0.0054 0.0078 0.0059 0.0049 0.0096 0.0115 0.006 0.0043 0.014 0.0045 0.0068 0.0019 0.0071 0.013 0.0029 0.0108 0.003 0.0039 0.0013 0.0072 0.0072 0.0052 0.006 0.015 0.0037 0.0029 0.0048 DEA 0.0048 0.0009 0.0018 0.0028 0.001 0.0058 0.0025 0.0005 0.0018 0.0015 0.0022 0.0043 0.0018 0.0011 0.002 0.0012 0.0029 0.0012 0.0011 0.0013 0.0016 0.001 0.0011 0.0016 0.0003 0.0019 0.0008 0.002 0.002 0.0013 0.0007 0.0019 0.0005 0.0021 0.0003 0.0042 0.0031 0.007 0 0.0027 0.0023 0.0045 0.0009 0.0014 0.003 0.0048 0.0041 0.0012 0.0034 0.0032 0.0007 0.0025 0.0043 0.0021 0.0047 0.0021 0.0009 0.0048 0.001 0.0018 0.004 0.0009 0.0087 0.003 0.0003 0.0023 DHEA 0.0116 0.0033 0.0106 0.0136 0.0045 0.0218 0.0039 0.0027 0.0104 0.0067 0.0094 0.0102 0.0066 0.0053 0.0127 0.0041 0.0109 0.0067 0.0021 0.0071 0.0049 0.0047 0.0027 0.004 0.002 0.0091 0.0049 0.0077 0.0042 0.0043 0.0041 0.0121 0.0059 0.0082 0.0029 0.0127 0.0058 0.0129 0.0059 0.0063 0.01 0.009 0.0071 0.0069 0.0162 0.0107 0.0103 0.0062 0.0153 0.0075 0.01 0.0062 0.0078 0.0267 0.0029 0.0182 0.0074 0.0048 0.0014 0.0081 0.0133 0.006 0.0049 0.0201 0.0145 0.0043 0.0073 PGF2a EA 0.0007 0.001 0.0011 0.0004 0.0016 15-HETE EA 0.0001 0 0.0003 0.0003 0.0004 0.0001 0 0.0001 0 NO-Gly 0.005 0.002 0.002 0.0059 0.0039 0.0044 0.0023 0.0015 0.0027 0.0037 0.0051 0.0025 0.0025 0.0038 0.0044 0.0021 0.0055 0.0025 0.0025 0.0026 0.002 0.0032 0.0019 0.0031 0.0036 0.0032 0.0023 0.0057 0.0015 0.0036 0.002 0.0068 0.0026 0.0059 0.0018 0.0067 0.0072 0.0116 0.0063 0.0084 0.0074 0.0057 0.0028 0.0042 0.0146 0.007 0.0079 0.0063 0.0029 0.0048 0.0061 0.0089 0.0031 0.0061 0.0137 0.0057 0.0038 0.0054 0.0031 0.0438 0.0021 0.0051 0.0049 0.0204 0.0047 0.0109 0.0026 PGE3 PGF3a PGE2 1G PGF2a 1G PGE2 EA PGD2 EA 11(12)-EpETre EA NA-Gly LTE4 0.074 0.0998 0.0356 0.0283 0.0752 0.0161 0.0983 0.0958 0.0647 0.0073 0.0639 0.118 0.0043 0.131 0.0449 0.0106 0.0403 0.0299 0.042 0.0899 0.0333 0.0136 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 1-OG 2-OG 1-LG 2-LG 1-AG 2-AG PEA SEA OEA LEA aLEA Dihomo GLA EA AEA DEA DHEA PGF2a EA 15-HETE EA NO-Gly PGE3 PGF3a PGE2 1G PGF2a 1G PGE2 EA PGD2 EA 11(12)-EpETre EA NA-Gly LTE4 METABOLITES_END #END