The MAGL inhibitor JZL184 and the NSAID diclofenac sodium salt were purchased from Cayman Chemical (Ann Arbor, MI). The gastric acid secretagogue pentagastrin and the proton pump inhibitor omeprazole were purchased from Sigma Aldrich (St. Louis, MO). All compounds were dissolved in a vehicle consisting of ethanol, Cremophor (Sigma-Aldrich), and saline in a ratio of 1:1:18 parts, as described previously (Crowe et al., 2015 (link); Kinsey and Cole, 2013 (link)). All solutions were warmed to room temperature and administered per oral (p.o.), intraperitoneally (i.p.), or subcutaneously (s.c.) at a volume of 10 μl/g body mass. Diclofenac was administered p.o., consistent with clinical use in that the NSAID comes into direct contact with the lining of the stomach. omeprazole and JZL184 were administered i.p., and pentagastrin was administered s.c.
Diclofenac sodium salt
Manufactured by Cayman Chemical
Sourced in United States
Diclofenac sodium salt is a chemical compound used as a laboratory reagent. It is the sodium salt form of diclofenac, a non-steroidal anti-inflammatory drug (NSAID). This salt is commonly used in research and scientific applications, but its specific function or intended use is not provided in this factual description.
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Lab products found in correlation
4 hydroxy s mephenytoin, by LGC (3 mentions)
Coumarin, by Merck Group (3 mentions)
S mephenytoin, by Cayman Chemical (3 mentions)
Testosterone, by Cayman Chemical (3 mentions)
4 hydroxydiclofenac, by Cayman Chemical (3 mentions)
7 hydroxycoumarin, by Chem Service (3 mentions)
Cremophor, by Merck Group (2 mentions)
Jzl184, by Cayman Chemical (2 mentions)
Phenacetin, by Merck Group (2 mentions)
4 aminobenzoic acid, by Merck Group (2 mentions)
Chlorzoxazone, by Merck Group (2 mentions)
Dextromethorphan hydrobromide, by Merck Group (2 mentions)
6β hydroxytestosterone, by Merck Group (2 mentions)
Paclitaxel, by Cayman Chemical (2 mentions)
Benzydamine hydrochloride, by Merck Group (2 mentions)
Kynuramine hydrobromide, by Santa Cruz Biotechnology (2 mentions)
6 hydroxychlorzoxazone, by LGC (2 mentions)
1 hydroxymidazolam, by LGC (2 mentions)
7 hydroxycoumarin β d glucuronide sodium salt, by Merck Group (2 mentions)
Dextrorphan tartrate, by Cayman Chemical (2 mentions)
7 protocols using diclofenac sodium salt
1
Gastric Ulcer Inhibition Protocol
2
Pharmacological Modulation of Cannabinoid Signaling
3
Cell Lysis and COX Activity Assay
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(1) Cell Lysate Preparation. MH-S cells were seeded at 106 cells/mL in a 48-well microplate and allowed to adhere for 24 h and then treated with guttiferone E-loaded liposomes or diclofenac at a different concentration as well as LPS 1 μg/mL. After 24 h, cells were detached with TNE buffer (Tris 40 mM, NaCl 150 mM, and EDTA 1 mM) and washed with PBS (1x), then resuspended in 1 mL PBS (1x), transferred into a 1.5 mL tube, and centrifuged at 500 × g for 3 min. The pellet was then resuspended in 0.5 mL of lysis buffer with a protease inhibitor cocktail, vortexed, and incubated in the ice for 5 min. The obtained cell lysate was centrifuged at 12000 × g for 3 min, and the supernatant was collected and kept on the ice for the COX activity assay.
(2) COX Activity Assay. The fluorometric cyclooxygenase (COX) activity assay kit (Biovision) was used according to the manufacturer's instructions to examine the ability of the GEL to inhibit the COX-1/COX-2 isozyme activity. The assay includes COX-1 and COX-2 specific inhibitors to differentiate the activity of COX-1 and COX-2 as well as other peroxidases. Diclofenac sodium salt (Cayman Chemical Company) 5 μM was used as a standard drug, and appropriate controls were also included.
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(1) Cell Lysate Preparation. MH-S cells were seeded at 106 cells/mL in a 48-well microplate and allowed to adhere for 24 h and then treated with guttiferone E-loaded liposomes or diclofenac at a different concentration as well as LPS 1 μg/mL. After 24 h, cells were detached with TNE buffer (Tris 40 mM, NaCl 150 mM, and EDTA 1 mM) and washed with PBS (1x), then resuspended in 1 mL PBS (1x), transferred into a 1.5 mL tube, and centrifuged at 500 × g for 3 min. The pellet was then resuspended in 0.5 mL of lysis buffer with a protease inhibitor cocktail, vortexed, and incubated in the ice for 5 min. The obtained cell lysate was centrifuged at 12000 × g for 3 min, and the supernatant was collected and kept on the ice for the COX activity assay.
(2) COX Activity Assay. The fluorometric cyclooxygenase (COX) activity assay kit (Biovision) was used according to the manufacturer's instructions to examine the ability of the GEL to inhibit the COX-1/COX-2 isozyme activity. The assay includes COX-1 and COX-2 specific inhibitors to differentiate the activity of COX-1 and COX-2 as well as other peroxidases. Diclofenac sodium salt (Cayman Chemical Company) 5 μM was used as a standard drug, and appropriate controls were also included.
4
High-Throughput Cytochrome P450 Profiling
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Acetaminophen, astemizole, bupropion, N‐acetyl‐p‐aminobenzoic acid, 4‐aminobenzoic acid, benzydamine hydrochloride, chlorzoxazone, coumarin, dextromethorphan hydrobromide, 7‐Ethoxyresorufin, 7‐Hydroxycoumarin β‐D‐glucuronide sodium salt, 7‐Hydroxycoumarin sulfate potassium salt, 6β‐hydroxy testosterone, irinotecan, midazolam, phenacetin, resorufin, SN38, and sulfamethazine were purchased from Sigma Aldrich. Dextrorphan tartrate, diclofenac sodium salt, 4‐hydroxydiclofenac, hydroxy bupropion, S‐mephenytoin, 4‐hydroxyquinoline, paclitaxel, and testosterone were purchased from Cayman Chemical. 7‐hydroxycoumarin was purchased from Chem Service. Benzydamine N‐oxide, kynuramine hydrobromide, and N‐acetyl sulfamethazine were obtained from Santa Cruz Biotechnology. Carbazeran, 4‐hydroxyCarbazeran, 6‐hydroxychlorzoxazone, 6α‐hydroxy paclitaxel, 1′‐hydroxymidazolam, and 4‐hydroxy‐S‐mephenytoin were obtained from Toronto Research Chemicals.
5
Diclofenac Sodium Salt Extraction Protocol
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Distilled water was received from Milli-Q Quantum Tex (Merck KGaA, Darmstadt, Germany, 18.2 MΩ cm, 3 ppb total organic carbon). Diclofenac sodium salt was obtained from Cayman Chemical Company (Ann Arbor, MI, USA). Ethanol (≥96%, denatured) and PVDF flat-sheet membranes with a pore size of 0.45 µm were acquired from Carl Roth GmbH + Co.KG (Karlsruhe, Germany). Soda-lime petri (Steriplan, Duran Group) dishes were purchased from DWK Life Sciences GmbH (Wertheim, Germany).
6
Metabolism Assay for Pharmacological Compounds
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Astemizole, irinotecan hydrochloride, diclofenac sodium salt, 7-ethyl-10-hydroxycamptothecin (SN38), 4-hydroxydiclofenac, s-mephenytoin, and testosterone were purchased from Cayman Chemical (Ann Arbor, MI). 7-HydroxyCoumarin was purchased from Chem Service (West Chester, Pennsylvania). 7-HydroxyCoumarin sulfate potassium salt was obtained from Santa Cruz Biotechnology (Dallas, Texas). Coumarin, 6β-hydroxytestosterone, 7-HydroxyCoumarin β-D-glucuronide sodium salt, (2S, 3S)-hydroxy were purchased from Sigma-Aldrich (St. Louis, MO). Midazolam, 1'-hydroxyMidazolam, O-desmethyl astemizole, and 4-hydroxy-s-mephenytoin were obtained from Toronto Research Chemicals (Toronto, Canada).
7
Acquisition of Enzyme Substrates
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Dextrorphan tartrate, diclofenac sodium salt, 4-hydroxydiclofenac, S-mephenytoin, 4-hydroxyquinoline, paclitaxel, and testosterone were purchased from Cayman Chemical (Ann Arbor, MI). 7-Hydroxycoumarin was purchased from Chem Service (West Chester, PA). Benzydamine N-oxide, 7-Hydroxycoumarin sulfate potassium salt, kynuramine hydrobromide, and N-acetyl sulfamethazine were obtained from Santa Cruz Biotechnology (Dallas, TX). 4-Acetamidobenzoic acid, p-acetamidophenyl b-D-glucuronide sodium salt, 4-aminobenzoic acid, benzydamine hydrochloride, chlorzoxazone, coumarin, dextromethorphan hydrobromide, 6b-hydroxytestosterone, 7-Hydroxycoumarin b-D-glucuronide sodium salt, 7-ethoxycoumarin, paracetamol sulfate potassium, phenacetin, and sulfamethazine were purchased from Sigma-Aldrich (St. Louis, MO). Carbazeran, 4-hydroxyCarbazeran, 6-hydroxychlorzoxazone, 6a-hydroxypaclitaxel, acetaminophen glutathione disodium salt, midazolam, 19-hydroxymidazolam, and 4-hydroxy-S-mephenytoin were obtained from Toronto Research Chemicals (Toronto, ON, Canada). All other drug-metabolizing enzyme substrates were obtained from Sigma-Aldrich.
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