Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Apr 24;8(4):e61696.
doi: 10.1371/journal.pone.0061696. Print 2013.

Atomic Hydrogen Surrounded by Water Molecules, H(H2O)m, Modulates Basal and UV-induced Gene Expressions in Human Skin in Vivo

Affiliations
Free PMC article

Atomic Hydrogen Surrounded by Water Molecules, H(H2O)m, Modulates Basal and UV-induced Gene Expressions in Human Skin in Vivo

Mi Hee Shin et al. PLoS One. .
Free PMC article

Abstract

Recently, there has been much effort to find effective ingredients which can prevent or retard cutaneous skin aging after topical or systemic use. Here, we investigated the effects of the atomic hydrogen surrounded by water molecules, H(H2O)m, on acute UV-induced responses and as well as skin aging. Interestingly, we observed that H(H2O)m application to human skin prevented UV-induced erythema and DNA damage. And H(H2O)m significantly prevented UV-induced MMP-1, COX-2, IL-6 and IL-1β mRNA expressions in human skin in vivo. We found that H(H2O)m prevented UV-induced ROS generation and inhibited UV-induced MMP-1, COX-2 and IL-6 expressions, and UV-induced JNK and c-Jun phosphorylation in HaCaT cells. Next, we investigated the effects of H(H2O)m on intrinsically aged or photoaged skin of elderly subjects. In intrinsically aged skin, H(H2O)m application significantly reduced constitutive expressions of MMP-1, IL-6, and IL-1β mRNA. Additionally, H(H2O)m significantly increased procollagen mRNA and also decreased MMP-1 and IL-6 mRNA expressions in photoaged facial skin. These results demonstrated that local application of H(H2O)m may prevent UV-induced skin inflammation and can modulate intrinsic skin aging and photoaging processes. Therefore, we suggest that modifying the atmospheric gas environment within a room may be a new way to regulate skin functions or skin aging.

Conflict of interest statement

Competing Interests: The authors have the following interests. Raeeun Park, Hideo Nojima and Hyung-Chel Kim are employed by Samsung Electronics CO., LTD. Part of this study was supported by a research agreement with Samsung Electronics CO., LTD. Samsung Electronics CO. LTD. is named in a patent application for the use of SPi device (atmospheric pressure plasma device releasing atomic hydrogen) as the type of air cleaner (patent actual). There are no further patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

Figures

Figure 1
Figure 1. H(H2O)m prevents UV-induced erythema and thymidine dimers in young human skin in vivo.
Skin from young human buttocks was irradiated with UV light and then locally treated with or without H(H2O)m for 2 hr. Twenty-four hours after irradiation, erythema index was measured and then skin was biopsied. (A) The photographs of erythema are representative of the subjects. (B) Erythema-index measurements are shown as means ± SEM with scatter plots (n = 11). (C) Immunohistochemical staining was performed using anti-thymidine dimer antibody. The figures shown are representative of seven subjects. (D) Results are expressed as means ± SEM with scatter plots (n = 7), *** p<0.001 versus the control, ## p<0.01 versus UV-irradiated skin.
Figure 2
Figure 2. H(H2O)m prevents and UV-induced MMP-1, COX-2, IL-6 and IL-1β mRNA expression in young human skin in vivo.
Skin from young human buttocks was irradiated with UV light and then locally treated with or without H(H2O)m for 2 hr. Twenty-four hours after irradiation, and then skin was biopsied. (A) MMP-1, (B) COX-2, (C) IL-6 and (D) IL-1β mRNA expressions were determined by real time RT-PCR. Results are expressed as means ± SEM with scatter plots (n = 11), * p<0.05, ** p<0.01, *** p<0.001. (E) Immunohistochemical staining was performed using anti-human MMP-1 antibody. The figures shown are representative of eleven subjects.
Figure 3
Figure 3. H(H2O)m prevents UV-induced MMP-1, COX-2 and IL-6 mRNA expression in HaCaT cells.
(A) Cells were pre-treated with H(H2O)m for 15 min, then irradiated with UV (55 mJ/cm2) and post-treated with H(H2O)m for 15 min in PBS. After incubation for 48 hr with serum-free DMEM, MMP-1 expression was determined in culture media and COX-2 and β-actin expression were determined in cell lysates by Western blotting. The bands are representative of results from three independent experiments. Results are expressed as means ± SEM (n = 6). (B) Cells were treated with H(H2O)m and UV as described above. After incubation for 24 hr with serum free DMEM, MMP-1, COX-2 and IL-6 mRNA expressions were determined by real-time RT-PCR. Results are expressed as means ± SEM (n = 3), * p<0.05, ** p<0.01, *** p<0.001.
Figure 4
Figure 4. UV-induced MMP-1 expression is mediated by ERK and JNK activation but not by p38 activation in HaCaT cells.
(A) Cells were serum-starved for 24 hr, irradiated with UV light (55 mJ/cm2) and harvested at the indicated times. Total cell lysates were prepared and subjected to Western blotting using specific antibodies. The bands are representative of results from three independent experiments. (B) HaCaT cells were pretreated with U0126(U) [ERK (MEK1) inhibitor; 10 µM], SP600125(SP) [JNK inhibitor; 10 µM], and SB203580(SB) [p38 inhibitor; 10 µM] for 1 hr, and then irradiated with UV. Irradiated cells were cultured for 48 hr and MMP-1 expression was determined by Western blotting. All experiments were performed in triplicate. Values shown are means ± SEM (n = 4), * p<0.05 versus the control, # p<0.05 versus UV-irradiated cells.
Figure 5
Figure 5. UV-induced SEK1/JNK activation and c-Jun phosphorylation are inhibited by H(H2O)m treatment in HaCaT cells.
(A) Cells were pre-treated with H(H2O)m for 15 min, then irradiated with UV and post-treated with H(H2O)m for 15 min in PBS. After incubation for 30 min, total cell lysates were prepared. Western blotting were performed using phospho-specific (p−) and total (T−) JNK, c-Jun, SEK1 and ERK antibodies. Level of β-actin was used as loading control. The bands are representative of results from three independent experiments. (B) Results are expressed as means ± SEM (n = 3), ** p<0.01 versus the control, # p<0.05 versus UV-irradiated cells. (C) Intracellular phospho-c-Jun levels in cells were visualized using a fluorescent microscope and the images presented are representative of the fluorescence levels observed in three separate experiments.
Figure 6
Figure 6. H(H2O)m reduces UV-induced ROS production in HaCaT cells.
(A) Cells were pre-treated with H(H2O)m for 15 min, then irradiated with UV and post-treated with H(H2O)m for 15 min in PBS. Subsequently, cells were treated with DCFDA (25 µM, freshly diluted in pre-warmed DMEM) for 30 min. Cells were then assayed using a fluorescence reader. Values shown are means ± SEM (n = 8), *** p<0.001 versus the control, ### p<0.001 versus UV-treated cells. (B) Intracellular H2O2 levels in HaCaT cells were visualized after DCF staining. The fluorescence intensity was visualized using a fluorescent microscope, and the images presented are representative of the fluorescence levels observed in three separate experiments. To normalize cell number, DAPI was used as a fluorescent marker for the nucleus.
Figure 7
Figure 7. Basal expression of MMP-1, COX-2 and IL-6 are decreased by H(H2O)m treatment in intrinsically aged human buttock skin and H(H2O)m increases basal expression of type I procollagen expression in photoaged facial skin and also reduces the basal expressions of MMP-1 and IL-6.
(A) Aged human buttock skin was topically treated with or without H(H2O)m using a releasing device for 2 hr. MMP-1, COX-2 and IL-6 mRNA expressions were determined by real time RT-PCR. Results are shown as means ± SEM with scatter plots (n = 7), * p<0.05. (B) Immunohistochemical staining was performed using anti-human MMP-1 antibody. The figures shown are representative of seven subjects. (C) Aged human face skin was topically treated with or without H(H2O)m for 30 min a day for 4 days. Type I procollagen, MMP-1 and IL-6 mRNA expressions were determined by real time RT-PCR. Results are expressed as means ± SEM with scatter plots (n = 10), * p<0.05, *** p<0.001.

Similar articles

See all similar articles

Cited by 5 articles

References

    1. Smith WL, Meade EA, DeWitt DL (1994) Pharmacology of prostaglandin endoperoxide synthase isozymes-1 and -2. Ann N Y Acad Sci 714: 136–142. - PubMed
    1. Wlaschek M, Heinen G, Poswig A, Schwarz A, Krieg T, et al. (1994) UVA-induced autocrine stimulation of fibroblast-derived collagenase/MMP-1 by interrelated loops of interleukin-1 and interleukin-6. Photochem Photobiol 59: 550–556. - PubMed
    1. Kirnbauer R, Kock A, Neuner P, Forster E, Krutmann J, et al. (1991) Regulation of epidermal cell interleukin-6 production by UV light and corticosteroids. J Invest Dermatol 96: 484–489. - PubMed
    1. Konnikov N, Pincus SH, Dinarello CA (1989) Elevated plasma interleukin-1 levels in humans following ultraviolet light therapy for psoriasis. J Invest Dermatol 92: 235–239. - PubMed
    1. Ichihashi M, Ueda M, Budiyanto A, Bito T, Oka M, et al. (2003) UV-induced skin damage. Toxicology 189: 21–39. - PubMed

Publication types

MeSH terms

Grant support

This study was supported by a grant from the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2011-0029819), and research agreement with Samsung Electronics CO., LTD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Feedback