Hydrogen-Rich Water Prevents Dehydration-Induced Cellular Oxidative Stress and Cell Death in Human Skin Keratinocytes

Abstract

Hypohydration is linked to increased risk of a variety of diseases and can be life-threatening, especially in elderly populations. Dehydration induces cellular damage partially through the production of reactive oxygen species (ROS) in cells, tissues and organs. Hydrogen molecules are known to convert ROS to harmless water. Therefore, theoretically hydrogen-rich water (HW) might eliminate dehydration-induced ROS and reverse its harmful effects in cells. In this in vitro study, we demonstrated that air-drying for 5 min could induce ROS generation in both nucleus and cytoplasm of human keratinocytes HaCaT as quantified by CellROX® Green/Orange reagents (Thermo Fisher Scientific, Waltham, Massachusetts, U.S.), respectively. Conversely, when the air-drying time was increased to 10 and 20 min, HaCaT cells lost the ability to produce ROS. Scanning electron microscopic (SEM) images showed that 10 min air-drying could induce severe membrane damage in HaCaT cells. PrestoBlue assay showed that, when HaCaT cells were air-dried for 20 min, cell viability was decreased to 27.6% of the control cells 48 h later. However, once HaCaT cells were pretreated with HW-prepared media, dehydration-induced intracellular ROS, cell membrane damage and cell death were significantly reduced as compared with double distilled water (DDW) under the same conditions. In conclusion, our data suggested that HW can decrease dehydration-induced harmful effects in human cells partially through its antioxidant capacity.