The Effect of Szigetvár Medicinal Water on HaCaT Cells Exposed to Dithranol

Abstract

(1) Introduction: Topical dithranol is still commonly used today as an effective treatment for psoriasis. Dithranol treatment is often supplemented with balneotherapy, which has been shown to increase effectiveness and reduce side effects. The inorganic salts (sulfhide, selenium, zinc) are usually thought to be responsible for the effect. The antioxidant effect of the waters is thought to be behind the therapeutic effect, for which inorganic substances (sulfides, selenium, zinc) are thought to be responsible. The organic matter content of medicinal waters is also particularly important, as humic acids, which are often found in medicinal waters, have antioxidant effects. (2) Methods: In this short-term experiment, we aimed to test the possible protective effect of Szigetvár medicinal water and its organic matter isolate on HaCaT cells exposed to dithranol. Malondialdehyde levels were measured, and RT-qPCR was used to investigate the gene expression of selected cytokines relevant in the oxidative stress response (IL-6, IL-8, TNF-α, GM-CSF) and the expression of microRNA-21. (3) Results: Szigetvár medicinal water and the organic isolate prevented the increase in malondialdehyde levels caused by dithranol treatment. The cytokine gene expressions elevated by dithranol exposure were reduced by the treatment. (4) Conclusions: Szigetvár medicinal water and organic substances alone may have a protective effect on patients' healthy skin surfaces against dithranol damage. We also demonstrated that the organic compounds are also responsible for the protective effect.

Keywords: MDA; dithranol; medicinal water; organic substances; oxidative stress; pro-inflammatory cytokine; psoriasis; thermal water.

Figure 1

The sequence of the primers used in the experiment.

Figure 2

(a) Survival of HaCaT cells in different dilutions of Szigetvár organic matter isolate in 3 h. (b) Survival of HaCaT cell in different concentrations of Szigetvár medicinal water containing cell culture medium (SVNM). 6×–3000×–dilution of the isolate (Composition shown in Table 1) in 10–90% Szigetvár medicinal water content of different studied SVNMs.

Figure 3

Amount of MDA (nmol/sample) in HaCaT cells in different experimental settings (See Table 2). (* = p < 0.05).

Figure 4

Relative gene expression level of TNF-α in HaCaT cells treated according to the experimental protocol. (* = p < 0.05). DTH—dithranol; 120×, 600×, 1200×-dilution of the tested organic matter isolate (Composition shown in Table 1); SVNM—Cell culture medium with 90% Szigetvár medicinal water content.

Figure 5

Relative gene expression level of GM-CSF in HaCaT cells treated according to the experimental protocol. (* = p < 0.05). DTH—dithranol; 120×, 600×, 1200×-dilution of the tested organic matter isolate (Composition shown in Table 1); SVNM—Cell culture medium with 90% Szigetvár medicinal water content.

Figure 6

Relative gene expression level of IL-6 in HaCaT cells treated according to the experimental protocol. (* = p < 0.05). DTH—dithranol; 120×, 600×, 1200×—dilution of the tested organic matter isolate (Composition shown in Table 1.); SVNM—Cell culture medium with 90% Szigetvár medicinal water content.

Figure 7

Relative gene expression level of IL-8 in HaCaT cells treated according to the experimental protocol. (* = p < 0.05). DTH—dithranol; 120×, 600×, 1200×-dilution of the tested organic matter isolate (Composition shown in Table 1); SVNM—Cell culture medium with 90% Szigetvár medicinal water content.

Figure 8

Relative gene expression level of miR-21 in HaCaT cells treated according to the experimental protocol. (* = p < 0.05). DTH—dithranol; 120×, 600×, 1200×-dilution of the tested organic matter isolate (Composition shown in Table 1); SVNM—Cell culture medium with 90% Szigetvár medicinal water content.