Figure 1.

Plasma from RA patients induced increased ROS production and reduced NO synthesis compared to healthy volunteers. (a) ECV-304 cells were cultured with 2.5 μM DCF for ROS and 5 μM DAF-2DA for 30 min at 37 °C 5% CO₂ in 199 media. The baseline for fluorescence intensity was determined using ECV-304 cells without stimulation. The positive fluorescence signal was obtained using 10 mM H₂O₂ stimulation for intracellular ROS and 10 μM histamine stimulation for intracellular NO. (b) ROS production and (c) NO synthesis was measured in ECV-304 cells cultured with plasma from RA patients and healthy volunteers after 12 h of incubation. Fluorescence intensity was measured using a microplate reader at emission 540 nm (excitation 485 nm) and confirmed by fluorescent microscope. (d) Protein concentration of hs-CRP, IL-6 and sVCAM-1 in plasma from RA patients and healthy controls was measured using a chemiluminescent immunometric solid phase assay and ELISA. Mann-Whitney test was used to compare RA patients and healthy controls, *** P < 0.0005 and **** P < 0.0001 was considered significant.