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, 20 (9), 16388-403

Gallic Acid Is the Major Active Component of Cortex Moutan in Inhibiting Immune Maturation of Human Monocyte-Derived Dendritic Cells

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Gallic Acid Is the Major Active Component of Cortex Moutan in Inhibiting Immune Maturation of Human Monocyte-Derived Dendritic Cells

Ben Chung Lap Chan et al. Molecules.

Abstract

Atopic dermatitis (AD) is a widely prevalent and chronically relapsing inflammatory skin disease. Penta Herbs Formula (PHF) is efficacious in improving the quality of life and reducing topical corticosteroid used in children with AD and one of the active herbs it contains is Cortex Moutan. Recent studies showed that altered functions of dendritic cells (DC) were observed in atopic individuals, suggesting that DC might play a major role in the generation and maintenance of inflammation by their production of pro-inflammatory cytokines. Hence, the aims of the present study were to identify the major active component(s) of Cortex Moutan, which might inhibit DC functions and to investigate their possible interactions with conventional corticosteroid on inhibiting the development of DC from monocytes. Monocyte-derived dendritic cells (moDC) culture model coupled with the high-speed counter-current chromatography (HSCCC), high pressure liquid chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry (LCMS) analyses were used. Gallic acid was the major active component from Cortex Moutan which could dose dependently inhibit interleukin (IL)-12 p40 and the functional cluster of differentiation (CD) surface markers CD40, CD80, CD83 and CD86 expression from cytokine cocktail-activated moDC. Gallic acid could also lower the concentration of hydrocortisone required to inhibit the activation of DC.

Keywords: Cortex Moutan; Penta Herb Formula; atopic dermatitis; dendritic cells; gallic acid; high-speed counter-current chromatography.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
High-Speed Counter-Current Chromatography (HSCCC) of Cortex Moutan. (a) The percentage of total weight of 30 HSCCC fractions; (b) The effects of the 30 fractions and Cortex Moutan aqueous extract (CM) on IL-12p40 production in monocyte-derived dendritic cells. Values represent mean ± SEM (n = 3). Fractions (200 µg/mL) were added to DC and cultured for 48 h.
Figure 2
Figure 2
HPLC profiles of bioactive markers (a) gallic acid; (b) paeoniflorin and (c) fractions 2, 3, 4 and 5 of Cortex Moutan HSCCC extract. Detection was performed at UV 274 nm. Representative retention time peaks of the bioactive markers are highlighted in circle. Fractions 4 and 5 showed retention time peak similar to that of gallic acid (highlighted in circle).
Figure 3
Figure 3
The effects of the active ingredients from Moutan Cortex on IL-12p40 production from human monocyte derived dendritic cells (moDC) (n = 3). (a) The Effects of fraction 5 (F5) and gallic acid were compared to the crude water extracts of Cortex Moutan and Penta Herb formula (PHF) on IL-12p40 production from human moDC, and hyrodocotroisone (2 μM) was used as positive control; (b) The effects of three active ingredients from Cortex Moutan: paeoniflorin, paeonol and gallic acid on IL-12p40 production from human moDC. * and *** indicate p < 0.05 and p < 0.001, respectively, when compared with the control level of the cytokine production or surface marker expression from cells without incubating with DC-inducing cytokine cocktail.
Figure 4
Figure 4
Effects of gallic acid on the surface expression of the functional markers on human monocyte-derived dendritic cells (moDC) (n = 4) Control (cells without adding DC-inducing cytokine cocktail); GA200: gallic acid (200 μg/mL); GA100: gallic acid (100 μg/mL); GA50: gallic acid (50 μg/mL); HC: hydrocortisone (2 μM). *, ** and *** indicate p < 0.05; p < 0.01 and p < 0.001, respectively, when compared the level of the cytokine production or surface marker expression from cells without incubated with DC-inducing cytokine cocktail (control). (a) CD40; (b) CD80; (c) CD83; (d) CD86; (e) CD11c; (f) HLA-DR; (g) CD14 and (h) IL12 p40.
Figure 5
Figure 5
Combined effects of gallic acid and hydrocortisone on the cytokine production from moDC (n = 10). The open bars represent the cells were added with gallic acid alone (0, 20 and 50 μg/mL) and the solid bars represent the cells added with gallic acid (0, 20 and 50 μg/mL) and hydrocortisone HC (0.2 μM). ** and *** indicate p < 0.01 and p < 0.001, respectively, when compared the control level of the cytokine production or surface marker expression from cells without incubating with DC-inducing cytokine cocktail. $ and $$$ indicate p < 0.05 and p < 0.001, respectively, when compared the control level of the cytokine production or surface marker expression cells with HC and without incubating with DC-inducing cytokine cocktail.
Figure 6
Figure 6
Combined effects of gallic acid and hydrocortisone on the expression of the functional markers from monocyte-derived dendritic cells (moDC) (n = 10). The open bars represent the cells were added with gallic acid alone (0, 20 and 50 μg/mL) and the solid bars represent the cells added with gallic acid (0, 20 and 50 μg/mL) and hydrocortisone HC (0.2 μM). * and ** indicate p < 0.05 and p < 0.01, respectively, when compared the control level of the cytokine production or surface marker expression from cells without incubating with DC-inducing cytokine cocktail. $$ indicate p < 0.01, respectively when compared the control level of the cytokine production or surface marker expression cells with HC and without incubating with DC-inducing cytokine cocktail.

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