Murine hemorrhagic peritonitis
This protocol is extracted from research article:
Resolution metabolomes activated by hypoxic environment
Sci Adv, Oct 23, 2019; DOI: 10.1126/sciadv.aax4895

All experimental procedures were approved by the Brigham and Women’s Hospital Institutional Animal Care and Use Committee (protocol no. 2016N000145) and complied with institutional and U.S. National Institutes of Health guidelines. Male Friend leukemia virus B (FVB) mice (6- to 8-week old) were given zymosan A (1 mg/0.5 ml; Sigma-Aldrich), thrombin (5 U/0.5 ml; Sigma-Aldrich), or both for 0, 12, or 24 hours using procedures reported in (20). Mice were then euthanized with isoflurane before peritoneal lavage was performed with 5.0 ml of ice-cold PBS without divalent cations. Lavages were subjected to LC-MS/MS for metabololipidomics analysis and flow cytometric analysis of neutrophil numbers, macrophage numbers, and macrophage efferocytosis of both neutrophils and erythrocytes (see “Flow cytometry” section above for antibodies and staining protocol). Resolution indices were calculated as in (65): Tmax (the time interval when PMN numbers reach maximum), T50 (the time interval when PMN numbers reach 50% of maximum), and Ri (the interval between Tmax and T50).

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