title: Bathing in carbon dioxide-enriched water alters protein expression in keratinocytes of skin tissue in rats.
abstract: Beneficial effects of balneotherapy using naturally occurring carbonated water (CO2enriched) 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 (CO2content 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.
authors.affiliation: Institute of Pathology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.|Department for Gastroenterology and Hepatology, Center for Internal Medicine, University Hospital of Essen, University Duisburg-Essen, Essen, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Kälsch
authors.affiliation: Institute of Pathology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.|Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
authors.forename: Leona L
authors.initials: LL
authors.lastname: Pott
authors.affiliation: Faculty of Health Sciences, Department of Rehabilitation Sciences, University of Tokyo Health Sciences, Tokyo, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Takeda
authors.affiliation: Mitsubishi Rayon Cleansui, Tokyo, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Kumamoto
authors.affiliation: Institute of Pathology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Dorothe
authors.initials: D
authors.lastname: Möllmann
authors.affiliation: Department for Gastroenterology and Hepatology, Center for Internal Medicine, University Hospital of Essen, University Duisburg-Essen, Essen, Germany.
authors.forename: Ali
authors.initials: A
authors.lastname: Canbay
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Sitek
authors.affiliation: Institute of Pathology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany. hideo.baba@uk-essen.de.
authors.forename: Hideo A
authors.initials: HA
authors.lastname: Baba
mesh_terms: D000818:Animals; D001452:Balneology; D061545:Carbonated Water; D049109:Cell Proliferation; D015603:Keratinocytes / Q000378:metabolism*; D008297:Male; D040901:Proteomics; D017208:Rats, Wistar; D012867:Skin / Q000378:metabolism
chemical_list: D061545:Carbonated Water
keywords: Balneotherapy; CO2; Carbonated water; Far upstream element binding protein 2 (FUBP2); Proteomics