Cromoglycate

In order to deplete neutrophils, mice received an i.v. injection of Anti-Ly6G (400 µg/mouse, 1A8 clone; eBiosciences) 12 h after the APAP challenge. We also performed different pharmacological intravenous interventions, always 12 h after the APAP challenge. To prevent neutrophil degranulation, mice were treated with: (i) cromoglycate (Sigma-Aldrich, St. Louis, MO, USA) in crescent doses of 5, 50 and 100 mg/kg; (ii) a protease inhibitor cocktail (Sigmafast^TM, St. Louis, MO, USA, EDTA-Free) in doses of 0.28, 1.4 and 2.8 mg/mouse; (iii) MPO inhibitor 4-aminobenzoic acid hydrazide (4-ABAH, Aldrich Chemistry, St. Louis, MO, USA), 80 mg/kg; (i.v.) matrix metalloproteinases inhibitors (MMP Inhibitor I, Calbiochem), 2 mg/kg; and (v) noncompetitive allosteric CXCR1/2 receptor antagonists (DF2156A, Dompé Farmaceutici SpA, L’Aquila AQ, Italy), 30 mg/kg. All inhibitors were tested alone before challenged with APAP and only doses that caused no injury were used in this study (data not shown).

FITC–dextran 150 kDa and dextran sulfate (DXS) 500 kDa were purchased from TdB Consultancy (Uppsala, Sweden). Heparin 4 kDa (enoxaparin EUROFARMA Versa, NC 154693) was kindly provided by PAS Mourão (Universidade Federal do Rio de Janeiro). HOE-140, B2R antagonist (B2RA) was purchased from Sigma (St. Louis, MO, USA). Bosentan was a donation from Actelion (Basel, Switzerland). FXII inhibitor (36 (link)) was a generous gift from Dr. Thomas Renné (Karolinska Institute). Cromoglycate, mepyramine, histamine, captopril were purchase from Sigma (St. Louis, MO, USA). PKSI-527 and PdSP15a were supplied by the co-authors (LJ) and (PA), respectively.