Effects of dextran sulfate, 4-t-butylcyclohexanol, pongamia oil and hesperidin methyl chalcone on inflammatory and vascular responses implicated in rosacea

Figures & data

Figure 1 Mean (pg/mL) and percentage inhibition of IL-1α and IL-8 expression (A), and RQ and percentage inhibition of KLK5 and MMP-9 mRNA expression (B) after incubation of NHEK with dextran sulfate for 24 hours in a rosacea environment. IKK inhibitor was used as a positive control.

Note: Data shown are the mean of three independent experiments. **P<0.01 vs control cells.
Abbreviations: RQ, relative quantity; MMP-9, matrix metalloproteinase 9; NHEK, normal human epidermal keratinocyte; IKK, I kappa B kinase.

Figure 2 Mean (pg/mL) and percentage inhibition of VEGF expression after incubation of keratinocytes with dextran sulfate for 24 hours in a rosacea environment.

Note: Data shown are the mean of three independent experiments. **P<0.01 vs control cells.
Abbreviation: VEGF, vascular endothelial growth factor.

Figure 3 Effect of dextran sulfate on pseudotube formation.

Notes: (A) Mean inhibition (%) of VEGF-induced pseudotube formation (mean of three experiments). Suramin 100 µM was used as a positive control. (B) Representative images of the effect of dextran sulfate on VEGF-induced pseudotube formation by endothelial cells. Endothelial cells were labeled with anti-collagen IV antibody-AF568. Magnification ×20. **p<0.01; ***p<0.001.
Abbreviation: VEGF, vascular endothelial growth factor.

Figure 4 Percentage TRPV1 inhibition after 30 minutes of stimulation by capsaicin and treatment with different concentrations of BCH.

Note: Data shown are the mean of three independent experiments.
Abbreviations: TRPV1, transient receptor potential vanilloid 1; BCH, 4-t-butylcyclohexanol.

Figure 5 Mean (pg/mL) and percentage inhibition of IL-8 expression after incubation of NHEK with BCH, pongamia oil, or BCH + pongamia oil, for 24 hours in a rosacea environment. IKK inhibitor was used as a positive control.

Notes: Data shown are the mean of three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs control cells.
Abbreviations: IKK, I kappa B kinase; BCH, 4-t-butylcyclohexanol; NHEK, normal human epidermal keratinocyte.

Figure 6 The RQ and percentage inhibition of IL-8, CXCL1, and CXCL6 mRNA expression after incubation of NHEK with BCH or pongamia oil for 24 hours in a rosacea environment. IKK inhibitor was used as a positive control.

Note: Data shown are the mean of three independent experiments.
Abbreviations: RQ, relative quantity; IKK, I kappa B kinase; BCH, 4-t-butylcyclohexanol; NHEK, normal human epidermal keratinocyte.

Figure 7 The activity of HMC in modulating vascular responses and IL-8 cytokine production after SP stimulation in human skin explants. Human skin explants were pre-incubated (or not) with HMC and then stimulated with SP for 24 hours. L-703,606 oxalate (10 µM), an NK1 inhibitor, was used as a positive control.

Notes: (A) Representative images from H&E coloration of the effect of the test compounds on vessel dilation: normal morphology of capillary vessels is denoted by an asterisk, dilated capillary vessels are marked by an arrow; SP induced a clear dilation of the capillary vessels, while the addition of NK1 inhibitor prevented vascular dilation. HMC prevented SP-induced vessels dilation. (B) CD34+ endothelial cells immunostaining (arrows); CD34-labeling of endothelial cells surrounded by capillary vessels revealed alteration of vessels by SP and normal morphology with both NK1 inhibitor and HMC. Magnification ×40. (C) Proportion of dilated vessel capillaries and percentage inhibition of dilation with the test compounds. (D) Mean (µm²) and percentage inhibition of vessels surface. (E) Mean (pg/mL) and percentage inhibition of IL-8 expression. Data shown are the mean of eight independent experiments. *P<0.05 and **P<0.01 vs SP-stimulated explants. ^##P<0.01 vs control explants. Red bar indicates exposure to rosacea environment only.
Abbreviations: HMC, hesperidin methyl chalcone; SP, substance P.