Fermented Taiwanofungus camphoratus Extract Ameliorates Psoriasis-Associated Response in HaCaT Cells via Modulating NF-?B and mTOR Pathways

Abstract

Psoriasis is a chronic autoimmune disease, and until now, it remains an incurable disease. Therefore, the development of new drugs or agents that ameliorate the disease will have marketing potential. Taiwanofungus camphoratus (TC) is a specific fungus in Taiwan. It is demonstrated to have anticancer, anti-inflammation, and hepatoprotective effects. However, the effects of TC fermented extract on psoriasis are under investigation. In this research, we studied the ability of TC on antioxidative activity and the efficacy of TC on interleukin-17 (IL-17A)-induced intracellular oxidative stress, inflammation-relative, and proliferation-relative protein expression in human keratinocytes. The results of a DPPH radical scavenging assay, reducing power assay, and hydroxyl peroxide inhibition assay indicated that TC has a potent antioxidant ability. Furthermore, TC could reduce IL-17A-induced intracellular ROS generation and restore the NADPH level. In the investigation of pathogenesis, we discovered TC could regulate inflammatory and cell proliferation pathways via p-IKKα/p-p65 and p-mTOR/p-p70S6k signaling pathways in human keratinocytes. In conclusion, TC showed characteristics such as antioxidant, anti-inflammatory, and anti-psoriatic-associated responses. It is expected to be developed as a candidate for oxidative-stress-induced skin disorders or psoriasis treatment.

Figures

Figure 1

HPLC chromatograms of sesamin and…

Figure 1

HPLC chromatograms of sesamin and EK100 in TC and standards. ( a )…

Figure 1 HPLC chromatograms of sesamin and EK100 in TC and standards. (a) sesamin standard, (b) EK100 standard, and (c) sesamin and EK100 in TC.

Figure 2

TC scavenges the DPPH radical…

Figure 2

TC scavenges the DPPH radical in a dose-dependent manner. TC exhibited a dose-dependent…

Figure 2 TC scavenges the DPPH radical in a dose-dependent manner. TC exhibited a dose-dependent DPPH radical scavenging ability, with an EC50 value of 205.7 µg/mL. Ascorbic acid was used as a positive control. Significant differences between the treated groups and the negative control were evaluated. *** p < 0.001 compared to the control group.

Figure 3

TC displays a concentration-dependent reducing…

Figure 3

TC displays a concentration-dependent reducing power. TC exhibited a dose-dependent reducing power with…

Figure 3 TC displays a concentration-dependent reducing power. TC exhibited a dose-dependent reducing power with an EC50 value of 857.9 µg/mL. Ascorbic acid was used as a positive control. Significant differences between the treated groups and the negative control were evaluated. *** p < 0.001 compared to the control group.

Figure 4

TC scavenges H 2 O…

Figure 4

TC scavenges H 2 O 2 in a dose-dependent manner. ( a )…

Figure 4 TC scavenges H2O2 in a dose-dependent manner. (a) The standard curve of absorbance at 230 nm at different hydrogen peroxide concentrations. (b) The effect of TC on H2O2 inhibition. TC showed a dose-dependent effect on H2O2 inhibition, with an EC50 value of 488.1 µg/mL and BHA as a positive control. Significant differences between treated groups and the negative control were evaluated. *** p < 0.001 compared to the control group.

Figure 5

Effect of ( a )…

Figure 5

Effect of ( a ) TC and ( b ) IL-17A on cell…

Figure 5 Effect of (a) TC and (b) IL-17A on cell viability in human keratinocytes. Significant differences between treated groups were evaluated. ** p < 0.01; *** p < 0.001 compared to the control group.

Figure 6

Effect of TC on cell…

Figure 6

Effect of TC on cell viability in IL-17A-induced human keratinocytes. Significant differences between…

Figure 6 Effect of TC on cell viability in IL-17A-induced human keratinocytes. Significant differences between treated groups were evaluated. ## p < 0.01 compared to the control group. *** p < 0.001 compared to the IL-17A-induced group.

Figure 7

( a ) IL-17A-induced and…

Figure 7

( a ) IL-17A-induced and ( b ) TC inhibits IL-17A-induced intracellular oxidative…

Figure 7 (a) IL-17A-induced and (b) TC inhibits IL-17A-induced intracellular oxidative stress in human keratinocytes. IL-17A at a concentration of 50 and 100 ng/mL significantly induced ROS generation, and TC suppressed IL-17A-induced ROS production in a dose-dependent manner in human keratinocytes. Significant differences between treated groups were evaluated. ### p < 0.001 compared to the control group. *** p < 0.001 compared to the IL-17A-induced group.

Figure 8

TC restores basal NADPH levels…

Figure 8

TC restores basal NADPH levels in IL-17A-treated human keratinocytes. TC could restore reduced…

Figure 8 TC restores basal NADPH levels in IL-17A-treated human keratinocytes. TC could restore reduced NADPH levels upon IL-17A treatment in human keratinocytes.

Figure 9

TC suppresses IL-17A-induced cell proliferation…

Figure 9

TC suppresses IL-17A-induced cell proliferation in human keratinocytes via the p -mTOR/ p…

Figure 9 TC suppresses IL-17A-induced cell proliferation in human keratinocytes via the p-mTOR/p-p70S6k signaling pathway. TC could inhibit IL-17A-induced p-mTOR and p-p70S6k expression in human keratinocytes, thereby possibly suppressing excess activation of the cell proliferation pathway. Significant differences between treated groups were evaluated. ### p < 0.001 compared to the control group. * p < 0.05; ** p < 0.01; *** p < 0.001 compared to the IL-17A-induced group.

Figure 10

TC inhibits IL-17A-induced expression of…

Figure 10

TC inhibits IL-17A-induced expression of pro-inflammatory proteins related to the NF-κB signaling pathway…

Figure 10 TC inhibits IL-17A-induced expression of pro-inflammatory proteins related to the NF-κB signaling pathway in human keratinocytes. TC could inhibit IL-17A-induced NF-κB expression in human keratinocytes, thereby possibly suppressing inflammatory responses. Significant differences between treated groups were evaluated. ### p < 0.001 compared to the control group. * p < 0.05; ** p < 0.01; *** p < 0.001 compared to the IL-17A-induced group.

Figure 11

Effect of TC on IL-6…

Figure 11

Effect of TC on IL-6 and IL-8 levels in IL-17A-treated human keratinocytes. (…

Figure 11 Effect of TC on IL-6 and IL-8 levels in IL-17A-treated human keratinocytes. (a) IL-6 level. and (b) IL-8 level. TC did not significantly reduce IL-17A-induced IL-6 and IL-8 levels in human keratinocytes.

Figure 12

Schematic diagram illustrating the inhibitory…

Figure 12

Schematic diagram illustrating the inhibitory effects of TC on IL-17A-induced inflammation and proliferation…

Figure 12 Schematic diagram illustrating the inhibitory effects of TC on IL-17A-induced inflammation and proliferation in human keratinocytes. All figures (12)