Figure 7.

Specific effect of HMW-HA in RPMI 2650 cells stimulated with rhIL-17A. HMW-HA was incubated with HAdase 2.5 units/100 μg (60°C for 72 hrs plus 65°C for 10 min). The cells were preincubated with HMW-HA (100 μg/mL) or HAdase treated HMW-HA for 1 h and stimulated with rhIL-17A (20 ng/mL) for 6 hrs for ROS or 18 hrs for NOX-4 and IL-8. (a) ROS production was evaluated in the cells by flow cytometry. Representative flow cytometry is shown. The results were expressed as the mean ± SD of 3 separate experiments; (b-c) NOX-4 and IL-8 protein synthesis were evaluated in the cell lysates by western blot. The bars represent ratio of band intensity and β-actin of 3 separate experiments. Representative western blot is shown. ANOVA with Fisher's test correction was used for the analysis of the data. p < 0.05 was statistically significant.