Pge2 elisa

Urinary PGE₂ levels were measured by highly sensitive PGE₂ ELISA (Enzo). To inhibit the activity of prostaglandin synthase, 10 μg/ml indomethacin (Alfa Aesar) was added. PGE₂ ELISA was performed on diluted urine (1:2) and standards. Optical density was measured at 405 nm with a Synergy H1 plate reader (BioTek). PGE₂ concentration was calculated based on the standard curve. Creatinine level was measured in diluted urine (1:20) and standards with the Creatinine Urinary Detection kit (Thermo Fisher Scientific). Absorbance was read at 490 nm and creatinine concentration was calculated based upon the standard curve.

Croton oil, carrageenan, CFA, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), dimethyl sulphoxide (DMSO) and LPS (Escherichia coli serotype 055:B5) were obtained from Sigma-Aldrich (St. Louis, MO, USA). MPO, SOD, MDA, total protein and NO kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). PGE₂ ELISA (enzyme-linked immunosorbent assay) kits were obtained from Enzo Life Sciences Inc. (NY, USA). TNF-α, IL-1β, and IL-6 ELISA kits were purchased from Boster Bioengineering Limited Company (Wuhan, China). All the other reagents were of analytical grade. Ultrapure water was used for the experiments.