15 hydroxyeicosatetraenoic acid hete d8

Lipidomic analysis was performed by LC–MS system as reported with modifications^{5 (link)}. In brief, samples were obtained by using centrifugal columns (Monospin C18-AX, GL Sciences, Tokyo, Japan) after the addition of deuterium-labeled internal standard [15-hydroxyeicosatetraenoic acid (HETE)-d8, arachidonic acid-d8, and leukotriene D₄-d4; Cayman Chemical, Ann Arbor, MI, USA]. Fatty acid metabolites were analyzed by using a UPLC system (Aquity; Waters, Milford, MA, USA) coupled with a mass spectrometer (Orbitrap Elite; ThermoFisher Scientific, Waltham, MA, USA). UPLC separation was performed with a 1.7-mm, 1.0 × 150 mm Aquity UPLC BEH C18 column (Waters). Mass spectrometric analysis for quantification was based on the ion-trap MS2 detection method in anion mode. Data analysis was performed by using the software Xcalibur 2.2 (ThermoFisher Scientific).

PQS (commercial name as Xinyue capsules, 50 mg PQS/capsule, National medicine permit No. Z20030072) was provided by Yisheng Pharmaceutical Co., Ltd (Jilin, China, batch No.180102). PQS was shown consistent quality between different batches and six ginsenosides (Rb1, Rb2, Rc, Rd, Re and Rg1) were detected as the major active compounds of PQS by our previous high-performance liquid chromatography ultraviolet analysis (the chemical structure of these ginsenosides were shown in Fig. S1) [11 (link)]. ASA was purchased from BAYER (Beijing, China). CLP was purchased from Sanofi (Paris, France). Adenosine diphosphate (ADP) were purchased from Chrono-log Corporation (Havertown, Pennsylvania, USA). 2,3,5-triphenyltetrazolium chloride (TTC) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Internal standards for lipidomics analysis including 6-keto-PGF1α-d4, Thromboxane (TX) B₂-d4, PGF2a-d4, PGE₂-d4, PGD₂-d4, Leukotriene (LT) C₄-d5, LTB₄ d4, 15-hydroxyeicosatetraenoic acid (HETE) -d8, 12-HETE-d8, 5-HETE -d8, AA-d8 were purchased from Cayman Chemical (Ann Arbor, Michigan, USA). The liquid chromatography-mass spectrometer (LC-MS)-grade solvents, methyl alcohol, acetonitrile, formic acid and ethanol were purchased from Thermo Fisher Scientific (Waltham, Massachusetts, USA).