Celecoxib

T24 or RT4 cells were seeded in 6-well plates 2×10⁵ cells/well, treated with corresponding reagents, including metformin (0, 1 mM, 5 mM, 10 mM, 20 mM), Celecoxib (20 μM YUANYE BioTECH, Shanghai, China) and PGE2(10 μM, Sigma). Cell lysates were separated on SDS-PAGE followed by western blotting assay as described [26 (link)] with the following primary antibodies: Bcl-2 (1:1000 Epitomics), Cyclin D1 (1:1000 Fantibody, China), CK14 (1:400 Abcam), STAT3 (1:2000 Abcam), OCT3/4 (1:400 Santa Cruz Biotech-nology), COX2 (1:2000 Origene), p-STAT3 (Tyr705) (1:2000 Cell Signaling), AMPK (1:700 Proteintech), p-AMPK (1:1000 Cell Signaling) and β-actin (1:5000 Cell Signaling).

To producing an inflammatory pain model, complete Freund’s adjuvant (CFA, Sigma-Aldrich, 10 μL) was injected intraplantarly. Luteoloside (HPLC≥98%, Shanghai Yuan Ye Biotechnology Co., Ltd) were dissolved in 1% DMSO (Sigma-Aldrich), and the concentration was adjusted to dosages of 20, 40, and 80 mg/kg by intraperitoneal injection, respectively. Celecoxib (HPLC≥98%, Shanghai Yuan Ye Biotechnology Co., Ltd) was dissolved in saline and adjusted to dosages of 20 and 40 mg/kg by intraperitoneal injection. All the drugs were prepared just before use.