Hydrogen Water Drinking Exerts Antifatigue Effects in Chronic Forced Swimming Mice via Antioxidative and Anti-Inflammatory Activities

Abstract

Purpose: This study was performed to evaluate antifatigue effect of hydrogen water (HW) drinking in chronic forced exercise mice model.

Materials and methods: Twelve-week-old C57BL6 female mice were divided into nonstressed normal control (NC) group and stressed group: (purified water/PW-treated group and HW-treated group). Stressed groups were supplied with PW and HW, respectively, ad libitum and forced to swim for the stress induction every day for 4 consecutive weeks. Gross antifatigue effects of HW were assessed by swimming endurance capacity (once weekly for 4 wk), metabolic activities, and immune-redox activities. Metabolic activities such as blood glucose, lactate, glycogen, blood urea nitrogen (BUN), and lactate dehydrogenase (LDH) as well as immune-redox activities such as reactive oxygen species (ROS), nitric oxide (NO), glutathione peroxidase (GPx), catalase, and the related cytokines were evaluated to elucidate underlying mechanism. Blood glucose and lactate were measured at 0 wk (before swimming) and 4 wk (after swimming).

Results: HW group showed a higher swimming endurance capacity (p < 0.001) than NC and PW groups. Positive metabolic effects in HW group were revealed by the significant reduction of blood glucose, lactate, and BUN in serum after 4 wk (p < 0.01, resp.), as well as the significant increase of liver glycogen (p < 0.001) and serum LDH (p < 0.05) than PW group. In parallel, redox balance was represented by lower NO in serum (p < 0.01) and increased level of GPx in both serum and liver (p < 0.05) than PW group. In line, the decreased levels of serum TNF-α (p < 0.01), IL-6, IL-17, and liver IL-1β (p < 0.05) in HW group revealed positive cytokine profile compared to PW and NC group.

Conclusion: This study shows antifatigue effects of HW drinking in chronic forced swimming mice via metabolic coordination and immune-redox balance. In that context, drinking HW could be applied to the alternative and safety fluid remedy for chronic fatigue control.

Figure 1

Increasing rate (%) of body weight time-dependently and swimming endurance capacity. Body weight percent difference (a) and swimming endurance capacity (b). Data were expressed as mean ± SD, n = 7. ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. NC: normal control group, PW: stressed group treated with purified water, and HW: stressed group treated with hydrogen water.

Figure 2

Metabolic activities. Blood glucose (a) and blood lactate (b) levels between wk 0 and wk 4, BUN in serum (c) and liver lysate (d), glycogen in serum (e) and liver lysate (f), and LDH level in serum (g) and liver lysate (h). Data were expressed as mean ± SD, n = 7. ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. NC: normal control group, PW: stressed group treated with purified water, and HW: stressed group treated with hydrogen water.

Figure 3

ROS and NO production. ROS level in serum (a) and liver lysate (b) and NO level in serum (c) and liver lysate (d). Data were expressed as mean ± SD, n = 7. ^∗p < 0.05, ^∗∗p < 0.01. NC: normal control group, PW: stressed group treated with purified water, and HW: stressed group treated with hydrogen water.

Figure 4

Antioxidant enzyme activities. GPx activity in serum (a) and liver lysate (b) and catalase activity in serum (c) and liver lysate (d). Data were expressed as mean ± SD, n = 7. ^∗p < 0.05, NC: normal control group, PW: stressed group treated with purified water, and HW: stressed group treated with hydrogen water.

Figure 5

Inflammatory cytokine analysis. IL-1β level in serum (a) and liver lysate (b), IL-6 level in serum (c) and liver lysate (d), IL-17 level in serum (e) and liver lysate (f), IL-23 level in serum (g) and liver lysate (h), and TNF-α level in serum (i) and liver lysate (j). Data were expressed as mean ± SD, n = 7. ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. NC: normal control group, PW: stressed group treated with purified water, and HW: stressed group treated with hydrogen water.