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Bathing in carbon dioxide-enriched water alters protein expression in keratinocytes of skin tissue in rats

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Abstract

Beneficial effects of balneotherapy using naturally occurring carbonated water (CO2 enriched) have been known since the Middle Ages. Although this therapy is clinically applied for peripheral artery disease and skin disorder, the underlying mechanisms are not fully elucidated.

Under controlled conditions, rats were bathed in either CO2-enriched water (CO2 content 1200 mg/L) or tap water, both at 37 °C, for 10 min daily over 4 weeks. Proliferation activity was assessed by Ki67 immunohistochemistry of the epidermis of the abdomen. The capillary density was assessed by immunodetection of isolectin-positive cells. Using cryo-fixed abdominal skin epidermis, follicle cells and stroma tissue containing capillaries were separately isolated by means of laser microdissection and subjected to proteomic analysis using label-free technique. Differentially expressed proteins were validated by immunohistochemistry.

Proliferation activity of keratinocytes was not significantly different in the epidermis after bathing in CO2-enriched water, and also, capillary density did not change. Proteomic analysis revealed up to 36 significantly regulated proteins in the analyzed tissue. Based on the best expression profiles, ten proteins were selected for immunohistochemical validation. Only one protein, far upstream element binding protein 2 (FUBP2), was similarly downregulated in the epidermis after bathing in CO2-enriched water with both techniques. Low FUBP2 expression was associated with low c-Myc immune-expression in keratinocytes.

Long-term bathing in CO2-enriched water showed a cellular protein response of epithelial cells in the epidermis which was detectable by two different methods. However, differences in proliferation activity or capillary density were not detected in the normal skin.

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Acknowledgment

The authors thank Laura Malkus who provided expertise that greatly assisted the research.

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Authors and Affiliations

  1. Institute of Pathology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany

    Julia Kälsch, Leona L. Pott, Dorothe Möllmann & Hideo A. Baba

  2. Faculty of Health Sciences, Department of Rehabilitation Sciences, University of Tokyo Health Sciences, Tokyo, Japan

    Atsushi Takeda

  3. Mitsubishi Rayon Cleansui, Tokyo, Japan

    Hideo Kumamoto

  4. Department for Gastroenterology and Hepatology, Center for Internal Medicine, University Hospital of Essen, University Duisburg-Essen, Essen, Germany

    Julia Kälsch & Ali Canbay

  5. Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany

    Leona L. Pott & Barbara Sitek

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  1. Julia Kälsch
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  2. Leona L. Pott
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  3. Atsushi Takeda
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  6. Ali Canbay
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  7. Barbara Sitek
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  8. Hideo A. Baba
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Correspondence to Hideo A. Baba.

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Sources of funding

This work was supported by intramural research funds of the Medical Faculty of the University of Duisburg-Essen (IFORES) and by Mitsubishi Rayon Cleansui.

Conflict of interest

Hideo Kumamoto is an employee of the company marketing the CO2 enriching device (Mitsubishi Rayon Cleansui).

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Kälsch, J., Pott, L.L., Takeda, A. et al. Bathing in carbon dioxide-enriched water alters protein expression in keratinocytes of skin tissue in rats. Int J Biometeorol 61, 739–746 (2017). https://doi.org/10.1007/s00484-016-1252-6

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