Antagonizing Effects of Clematis apiifolia DC. Extract against Benzo[a]pyrene-Induced Damage to Human Keratinocytes

Figure 2

CAE decreases benzo[a]pyrene- (B[a]P-) induced production of reactive oxygen species (ROS) in HaCaT cells. HaCaT cells were incubated with CAE in the presence of B[a]P (15 μM) for 24 h and then subjected to fluorescence image analysis (a) and fluorescence intensity analysis (b). HaCaT cells were incubated with CAE in the presence of H₂O₂ (55 μM) for 24 h and then subjected to fluorescence intensity analysis (c). The data were confirmed by repeating three independent experiments. Data are expressed as the vs. B[a]P-treated control. (d, e) HaCaT cells were incubated with CAE in the presence of B[a]P (15 μM) for 24 h and then subjected to an ELISA for proinflammatory cytokines. The results were confirmed by repeating three independent experiments. Data are expressed as the vs. untreated control; ^o vs. B[a]P-treated control. (f–h) HaCaT cells were transfected with siRNA for the AhR gene or Nrf2 gene using the DharmaFECT® Duo transfection reagent. After 24 h, the cells were incubated with CAE (100 μg/ml) in the presence of B[a]P (15 μM) for 24 h and then were subjected to an ELISA for IL-8 (f, g) and Western blot analysis for AhR or Nrf2 (h). The results were confirmed by repeating three independent experiments. Data are expressed as the vs. B[a]P-treated control; ^o vs. B[a]P plus CAE-treated control. CAE: Clematis apiifolia DC.; NAC: N-acetyl cysteine.

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