Tumor necrosis factor α tnf α

The human synovial cell line SW982 was purchased from Pricella (Wuhan, China). The cells were cultured in DMEM containing 10% extra fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin at 37°C with 5% CO2 in air. The culture medium was changed once every 2–3 days, and the cells were passaged when the cell fusion reached 90% or more. Tumor necrosis factor-α (TNF-α) used for in vitro stimulation was purchased from Proteintech (Wuhan, China). The cells were stimulated with a certain concentration of TNF-α for a period of time and collected for subsequent experiments.

Gancaonin N was purchased from Chemfaces (Wuhan Chemfaces Biochemical Co., Ltd., Wuhan, China). LPS (Escherichia coli 055:B5) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies for COX-2, NF-κB p65, extracellular signal-regulated kinase (ERK), p-ERK, p38, p-p38, and Lamin B1 were purchased from Cell Signaling Technology (Beverly, MA, USA), interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) were purchased from Proteintech (Rosemont, IL, USA), PGE2 was purchased from Bioss (Woburn, MA, USA), and iNOS was purchased from R&D Systems Inc. (Minneapolis, MN, USA). β-Actin and secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).