Development of Alkaline Reduced Water Using High-Temperature-Roasted Mineral Salt and Its Antioxidative Effect in RAW 264.7 Murine Macrophage Cell Line

Abstract

Oxidative stress (OS) plays an important role in many diseases, and its excessive increase affects human health. Although the antioxidant effect of sea salt can be strengthened through special processing, it is scarcely studied. This study confirmed the antioxidative effect of high-temperature roasted mineral salt (HtRMS) produced through repeated roasting of sea salt at high temperature in a ceramic vessel. The dissolved HtRMS exhibited properties such as high alkalinity, rich mineral content, and a high concentration of hydrogen (H2). To detect the antioxidative effect of HtRMS, OS was induced in RAW 264.7 murine macrophage cells with hydrogen peroxide (H2O2) and lipopolysaccharide (LPS), and then treated with HtRMS solution at different concentrations (0.1, 1, and 10%). Cell viability, reactive oxygen species (ROS), nitric oxide (NO), and antioxidant enzymes such as catalase (CAT) and glutathione peroxidase (GPx), Ca2+, and mitogen-activated protein kinase (MAPK) pathway-related proteins (p-p38, p-JNK, and p-ERK) were measured. OS was significantly induced by treatment with H2O2 and LPS (p < 0.001). After treatment with HtRMS, cell viability and GPx activities significantly increased and ROS, NO, Ca2+, and CAT significantly decreased in a concentration-dependent manner compared to H2O2 and LPS-only groups, which was not observed in tap water (TW)-treated groups. Similarly, p-p38, p-JNK, and p-ERK levels significantly decreased in a concentration-dependent manner in HtRMS groups compared to both H2O2 and LPS-only groups; however, those in TW groups did not exhibit significant differences compared to H2O2 and LPS-only groups. In conclusion, our results suggest that HtRMS may have antioxidant potential by regulating the MAPK signaling pathway.