Fig. 1

Fetuin A in supernatants of HepG2 cell cultures treated with proinflammatory cytokines. Human hepatocellular carcinoma HepG2 cells were cultured at 37 °C with 5 % CO₂ in high-glucose DMEM medium containing 10 % fetal bovine serum, 100 units/mL penicillin, and 100 μg/mL streptomycin in 6-well plates at a density of 0.25 × 10⁶ cells/mL until about 70 % confluence and old medium was replaced with fresh medium. Cell monolayers were then treated with rhTNF-α (50 ng/mL), rhIL-1β (10 ng/mL), rhIFN-γ (50 ng/mL), rhMCP-1 (40 ng/mL), and rhIL-6 (100 ng/mL) and incubated at 37 °C for 24 h. Cell supernatants were collected and fetuin-A levels were measured using sandwich high-sensitivity ELISA (Human fetuin-A PicoKine^TM ELISA kit, Boster Biological Technology, USA) following the manufacturer’s instructions as described in Patients and Methods. Fetuin-A production (mean ± SEM) was found to be significantly suppressed in HepG2 cells treated with TNF-α (82.05 ± 1.16 ng/mL, P = 0.002), IL-1β (82.73 ± 1.45 ng/mL, P = 0.003), and IFN-γ (85.70 ± 1.93 ng/mL, P = 0.02) as compared with untreated control (95.73 ± 1.43 ng/mL). However, fetuin-A production in cells treated with MCP-1 (84.74 ± 5.02 ng/mL, P = 0.10) and IL-6 (91.66 ± 2.55 ng/mL, P = 0.24) differed non-sidnificantly from control. The representative data from three independent determinations are shown