Naproxen

Monoclonal antibodies (mAb) against HO-1 and phospho-p65^Ser536 were from Abcam (Cambridge, UK). E-selectin mAb 1.2B6, ICAM-1 mAb 6.5B5 and VCAM-1 mAb 1.4C3 were generated in house. CD31 mAb P2B1 was generated in house from a clone purchased from Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA. The following antibodies were purchased from Cell Signaling (Danvers, MA) AMPKα, phospho-AMPKα^Thr172, CREB, phospho-CREB^Ser133. Antibodies against p65 and Nrf2 were from Santa Cruz Biotechnology (Heidelberg, Germany), α-tubulin Sigma-Aldrich (Poole, UK) and GAPDH Millipore (Watford, UK). 2,5-Dimethyl-Celecoxib was synthesized as described^{58 (link)}. Celecoxib was purchased from Toronto Research Chemicals (Ontario, Canada), naproxen and ibuprofen from Tocris Bioscience (Bristol, UK). The concentrations of all three drugs used in vitro (Celecoxib 1–10 µM, naproxen and ibuprofen 1–100 μM) included concentrations that can be achieved therapeutically in humans^{30 (link),38 (link),59 (link),60 (link)}. 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) and Compound C were from Millipore, hemin and sulforaphane from Sigma-Aldrich, TNFα and IL-1β from R and D Systems (Abingdon, UK).

Naproxen (Tocris, 10 mg/kg), codeine (St. Martins Pharmacy, 7 or 30 mg/kg), sumatriptan (Tocris, 10 mg/kg), and gabapentin (Sigma, 30 mg/kg) were dissolved in physiological saline (Braun) and administered via a single i.p. injection. Phenytoin (Sigma, 15 mg/kg) and aprepitant (MedChemExpress, 20 mg/kg) were dissolved in PEG-400 (Sigma) and delivered via i.p. injections. Drug injection schedules are detailed in Figure 1A: Naproxen, sumatriptan, and aprepitant were administered once on the last testing day. Codeine was administered at the last testing day twice (1 h before testing and when testing was concluded–before the night cycle). Phenytoin was administered once daily over a 15 day period, starting with the first injection of oxaliplatin. gabapentin was injected once per day over a period of 5 consecutive days starting 2 days after CCI-surgery.