Figure 7.

The effect of paclitaxel modulating inflammation, oxidative stress, apoptosis angiogenesis, and p-NF-κB is reduced in siRNA-TLR4 transiently transfected keratinocytes. Normal human epidermal keratinocytes (NHEK) were transiently transfected with control siRNA (-) or siRNA-TLR-4 and incubated for 24 h with paclitaxel 3 µM. (A–C) IL-1α, IL-6 and IL-8 supernatant levels were measured by ELISA. (D) Reactive oxygen species (ROS) were measured using H₂DCF-DA assay in NHEK stimulated with paclitaxel for 4 h. (E–J) The expression of SOD1, NOX4, Nrf2, BCL2, eNOS, and VEGF was measured by real-time PCR. Data are expressed as 2^−ΔCt. (K) NHEK cells were incubated for 1 h with paclitaxel concentrations. NF-κB and p-NF-κB protein levels were analyzed by Western blotting. Quantification was performed by densitometry and normalized to NF-κB/β-actin. Results are expressed as mean ± standard deviation of three independent experiments (n = 3). Multiple comparisons analysis of variance (ANOVA) was followed by the post hoc Bonferroni test. * p < 0.05 vs. siRNA (-) Control. # p < 0.05 vs. siRNA (-) PTX 3 µM. PTX: paclitaxel.