Antioxidant

Humans

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Pilot study: Effects of drinking hydrogen-rich water on muscle fatigue caused by acute exercise in elite athletes

Aoki, K.; Nakao, A.; Adachi, T.; Matsui, Y.; Miyakawa, S.
Source: Medical Gas Research, Volume 2, Article 12 (2012)
https://doi.org/10.1186/2045-9912-2-12


Background

Muscle contraction during short bouts of high-intensity exercise induces oxidative stress, which may contribute to overtraining symptoms, such as increased fatigue and muscular microinjury or inflammation. Previous studies have shown hydrogen’s antioxidant properties, prompting an investigation into the effect of hydrogen-rich water (HW) on oxidative stress and exercise-induced muscle fatigue.


Methodology

Ten male elite soccer players (mean age 20.9 ± 1.3 years) participated in this double-blind, placebo-controlled, crossover study. Each participant was tested twice, receiving either hydrogen-rich water (HW) or placebo water (PW) one week apart.

Participants performed 30 minutes of cycling at 75% of VO₂ max, followed by 100 repetitions of maximal isokinetic knee extension to assess peak torque and muscle performance. Blood samples were collected to measure oxidative stress markers (d-ROMs and BAP), creatine kinase (CK), and lactate levels.


Results

  • Blood lactate levels increased significantly during exercise with placebo water, but did not rise with hydrogen-rich water.

  • Peak torque during maximal knee extensions declined significantly in the placebo group, indicating fatigue. In contrast, the hydrogen group maintained torque in the early phase of the task.

  • No significant changes in oxidative stress markers or creatine kinase were observed post-exercise in either group.


Conclusion

Hydrogen-rich water consumed before intense exercise appears to reduce blood lactate accumulation and preserve muscle function during exhaustive performance. While the mechanisms and broader benefits require validation in larger studies, these preliminary findings suggest hydrogen-rich water could be an effective hydration strategy for athletes.

Effect of Hydrogen-Rich Water on Oxidative Stress, Liver Function, and Viral Load in Patients with Chronic Hepatitis B

Xia, C.X.; Liu, W.W.; Zeng, D.X.; Zhu, L.Y.; Sun, X.L.; Sun, X.J.
Source: Clinical and Translational Science, Volume 6, Issue 5 (2013)
https://doi.org/10.1111/cts.12076


Objective

To evaluate the effects of hydrogen-rich water (HRW) on oxidative stress, liver function, and HBV DNA levels in patients with chronic hepatitis B (CHB).


Methodology

Sixty patients with CHB were randomly divided into two groups:

  • A routine treatment group

  • A hydrogen treatment group, who consumed 1200–1800 mL of HRW orally, twice daily for six consecutive weeks.

Serum oxidative stress markers, liver function tests, and HBV DNA levels were measured before and after treatment. A group of 30 healthy individuals served as a control group.


Results

  • Compared to healthy controls, CHB patients showed elevated oxidative stress and impaired liver function.

  • After six weeks:

    • Oxidative stress remained unchanged in the routine treatment group but significantly improved in the hydrogen group.

    • Both groups showed notable improvement in liver function and significant reductions in HBV DNA levels.

  • Although there was a significant difference in oxidative stress between the two treatment groups, improvements in liver function and HBV DNA levels were comparable, both showing a positive trend.


Conclusion

Hydrogen-rich water markedly reduces oxidative stress in patients with chronic hepatitis B. While preliminary findings indicate possible improvements in liver function and viral load, further long-term studies are needed to validate HRW’s therapeutic potential in CHB management.

Consumption of water containing a high concentration of molecular hydrogen reduces oxidative stress and disease activity in patients with rheumatoid arthritis: an open-label pilot stud

Ishibashi, T.; Sato, B.; Rikitake, M.; Seo, T.; Kurokawa, R.; Hara, Y.; Naritomi, Y.; Hara, H.; Nagao, T.
Source: Medical Gas Research, Volume 2, Article 27 (2012)
https://doi.org/10.1186/2045-9912-2-27


Background

Rheumatoid arthritis (RA) is a chronic inflammatory disease marked by bone and cartilage destruction. While the exact cause remains unknown, hydroxyl radicals are believed to contribute to RA pathogenesis. Molecular hydrogen (H₂) has recently been identified as a selective scavenger of hydroxyl radicals, and methods have been developed to produce high-concentration hydrogen water. It is hypothesized that H₂-enriched water may complement conventional RA therapy by reducing oxidative stress.


Methodology

  • Participants: 20 patients with RA

  • Intervention: Daily consumption of 530 mL of water containing 4–5 ppm molecular hydrogen for 4 weeks

  • Washout period: 4 weeks

  • Re-administration: Same dose for an additional 4 weeks

  • Assessments:

    • Urinary 8-hydroxydeoxyguanosine (8-OHdG) as a marker of oxidative stress

    • Disease activity score (DAS28) using C-reactive protein (CRP)


Results

  • Drinking high-concentration H₂ water resulted in greater in vivo H₂ levels than H₂-saturated water (1.6 ppm).

  • Urinary 8-OHdG levels decreased significantly by an average of 14.3% (p < 0.01) after the first 4 weeks.

  • DAS28 score dropped from 3.83 to 3.02 (p < 0.01) during the same period.

  • After the washout, both 8-OHdG and DAS28 remained below baseline but showed slight increases.

  • In the second intake period, DAS28 dropped further from 2.83 to 2.26 (p < 0.01).

  • No further reduction in 8-OHdG was seen, but it stayed below the initial value.

  • All five early-stage RA patients (disease duration <12 months) who were ACPA-negative reached remission, and four became symptom-free by the study’s end.


Conclusion

These findings suggest that molecular hydrogen, as a hydroxyl radical scavenger, effectively reduces oxidative stress in RA patients. Clinical symptoms improved significantly with high-concentration H₂ water, particularly in patients with early RA.

Effects of Drinking Hydrogen-Rich Water on the Quality of Life of Patients Treated with Radiotherapy for Liver Tumors

Kang, K.M.; Kang, Y.N.; Choi, I.B.; Gu, Y.; Kawamura, T.; Toyoda, Y.; Nakao, A.
Source: Medical Gas Research, Volume 1, Article number 11 (2011)
https://doi.org/10.1186/2045-9912-1-11


Background

Cancer patients undergoing radiotherapy often experience fatigue and reduced quality of life (QOL). These side effects are largely attributed to oxidative stress and inflammation, caused by the generation of reactive oxygen species (ROS) during radiation exposure. Hydrogen, due to its antioxidant and anti-inflammatory properties, has been proposed as a therapeutic medical gas. This study explored whether hydrogen-rich water (HRW) could improve QOL in patients undergoing radiotherapy for malignant liver tumors.


Methodology

  • Design: Randomized, placebo-controlled clinical study

  • Participants: 49 patients receiving radiotherapy for malignant liver tumors

  • Intervention: Patients consumed HRW (0.55–0.65 mM H₂, prepared by magnesium stick reaction) daily for 6 weeks

  • Assessments:

    • QOL: Evaluated using the Korean version of the EORTC QLQ-C30 questionnaire

    • Oxidative stress: Measured by derivatives of reactive oxygen metabolites (d-ROMs)

    • Antioxidant capacity: Assessed via biological antioxidant potential (BAP)


Results

  • HRW consumption significantly reduced ROS levels in peripheral blood during radiotherapy.

  • The oxidative balance (BAP) was maintained in the HRW group but not in the placebo group.

  • QOL scores improved significantly in the HRW group compared to placebo during radiotherapy.

  • Importantly, there was no difference in tumor response to radiotherapy between the two groups, indicating that HRW did not interfere with anticancer effects.


Conclusion

Daily intake of hydrogen-rich water may be a novel therapeutic strategy to improve quality of life in patients exposed to radiation. It appears to attenuate oxidative stress-induced biological reactions without compromising the efficacy of radiotherapy.

Hydrogen-Rich Water for Improvements of Mood, Anxiety, and Autonomic Nerve Function in Daily Life

Mizuno, K.; Sasaki, A.T.; Ebisu, K.; Tajima, K.; Kajimoto, O.; Nojima, J.; Kuratsune, H.; Hori, H.; Watanabe, Y.
Source: Medical Gas Research, Volume 7, Issue 4 (2017), pp. 247–255
https://doi.org/10.4103/2045-9912.222448


Background

Everyone seeks health and a vibrant life. Improving quality of life (QOL), maintaining well-being, and preventing disease are crucial goals. Chronic oxidative stress and inflammation negatively affect the central nervous system, which can impair QOL. Even in healthy individuals, aging, work-related stress, and cognitive load over a few hours can increase oxidative stress. This suggests that preventing oxidative stress accumulation from daily stress and work may support overall QOL and slow aging-related decline.

Molecular hydrogen (H₂) has known antioxidant and anti-inflammatory properties and may therefore help improve QOL.


Objective

To evaluate the effects of drinking hydrogen-rich water (HRW) on QOL in healthy adult volunteers using psychophysiological assessments, including:

  • Psychological questionnaires

  • Tests of autonomic nerve function

  • Tests of cognitive function


Methods

  • Design: Double-blind, placebo-controlled, randomized, crossover trial

  • Participants: 26 healthy adults (13 women, 13 men; average age: 34.4 ± 9.9 years)

  • Intervention:

    • HRW group: 600 mL/day of hydrogen-rich water

    • PLW group: 600 mL/day of placebo water

  • Duration: Each participant underwent a 4-week treatment period with HRW or PLW, followed by a crossover.


Results

  • The post/pre-treatment ratios for K6 psychological distress scores and sympathetic nerve activity at rest were significantly lower after HRW intake compared to placebo.

  • This indicates improvements in mood, anxiety, and autonomic nervous system balance.


Conclusion

Drinking hydrogen-rich water may contribute to improved quality of life by supporting central nervous system functions, particularly in areas related to mood regulation, anxiety reduction, and autonomic nerve modulation.

Effects of the Long-Term Consumption of Hydrogen-Rich Water on the Antioxidant Activity and the Gut Flora in Female Juvenile Soccer Players from Suzhou, China

Sha, J.B.; Zhang, S.S.; Lu, Y.M.; Gong, W.J.; Jiang, X.P.; Wang, J.J.; Qiao, T.L.; Zhang, H.H.; Zhao, M.Q.; Wang, D.P.; Xia, H.; Li, Z.W.; Chen, J.L.; Zhang, L.; Zhang, C.G.
Source: Medical Gas Research, Volume 8, Issue 4 (2019), pp. 135–143
https://doi.org/10.4103/2045-9912.248263


Background

High levels of physical exertion during football training and competition inevitably lead to fatigue. An increasing number of studies confirm the association between the generation and removal of free radicals, exercise-induced fatigue, and injury. Recently, hydrogen has been identified as a novel selective antioxidant with potential beneficial applications in sports.


Objective

To evaluate the effect of two months of hydrogen-rich water (HRW) consumption on the gut microbiota of female juvenile soccer players from Suzhou, China.


Results

Based on ELISA and 16S rDNA sequencing of fecal samples, two months of HRW consumption led to:

  • Significant reductions in serum levels of:

    • Malondialdehyde (MDA)

    • Interleukin-1 (IL-1)

    • Interleukin-6 (IL-6)

    • Tumor necrosis factor-alpha (TNF-α)

  • Significant increases in:

    • Superoxide dismutase (SOD) levels

    • Total antioxidant capacity (T-AOC)

    • Hemoglobin in whole blood

Additionally, HRW improved the diversity and abundance of gut microbiota:

  • After two months of intervention, Shannon, Simpson, ACE, and Chao1 diversity indices were higher than pre-intervention levels and those of the control group.

  • However, prior to the intervention, there were already differences in microbiota composition between the two groups. During the trial period, no major changes were observed in the overall composition.


Conclusion

Two-month consumption of hydrogen-rich water may contribute to the modulation of gut flora in athletes, likely due to its selective antioxidant and anti-inflammatory properties.

Effects of concomitant use of hydrogen water and photobiomodulation on Parkinson disease: A pilot study

Hong, C.T.; Hu, C.J.; Lin, H.Y.; Wu, D.
Source: Medicine (Baltimore), Volume 100, Article number 4 (2021)
https://doi.org/10.1097/MD.0000000000024191


Background

Parkinson’s disease (PD), the second most common neurodegenerative disorder, currently has no curative or disease-modifying treatments — only symptomatic therapies. Oxidative stress and mitochondrial dysfunction play key roles in the pathophysiology of PD.

Animal studies have shown that photobiomodulation (PBM) can enhance mitochondrial function and increase ATP production, potentially alleviating PD symptoms. However, PBM may also lead to elevated levels of reactive oxygen species (ROS).

Molecular hydrogen (H₂) is a potent antioxidant that may counteract ROS effects. When PBM targets the brainstem, it can stimulate neuronal activity, while H₂ can neutralize excess ROS generated by PBM. This study aimed to evaluate the safety and efficacy of combined PBM and H₂ therapy in PD patients.


Methods

  • Participants: 18 patients with PD, aged 30–80 years, Hoehn and Yahr stages II–III.

  • Intervention: All received daily PBM + hydrogen water therapy for 2 weeks.

  • Assessment: Monitored for adverse events and changes in Unified Parkinson Disease Rating Scale (UPDRS) scores.


Results

  • UPDRS scores significantly decreased starting from week 1, with improvement continuing until the end of the therapy.

  • No adverse effects were reported.

  • One week after therapy cessation, UPDRS scores slightly increased, but remained significantly improved compared to baseline.


Conclusion

This pilot study demonstrated that the combination of PBM and hydrogen water is safe and effectively reduces disease severity in PD patients. A larger-scale clinical trial is planned to further investigate the therapeutic effects of PBM + H₂ on Parkinson’s disease.

Drinking Hydrogen-Rich Water for 4 Weeks Positively Affects Serum Antioxidant Enzymes in Healthy Men: A Pilot Study

Trivic, T.; Vojnovic, M.; Drid, P.; Ostojic, S.M.
Source: Current Topics in Nutraceutical Research, Volume 15, Issue 1 (2017)


Background

Molecular hydrogen is an experimental therapeutic agent in biomedicine, but its effects on serum antioxidant markers are still not well characterized.


Methodology

  • Participants: 16 healthy male volunteers (mean age: 24.6 ± 3.6 years; height: 183.3 ± 4.4 cm; weight: 83.5 ± 12.5 kg).

  • Design: Randomized, double-blind, placebo-controlled pilot study.

  • Intervention: Participants were assigned to receive either 300 ml/day of orally administered hydrogen-rich water (HRW) or tap water (placebo) for 4 weeks.

  • Measurements: Baseline and post-4-week levels of serum antioxidant enzymes.


Results

  • Significant improvements were observed in:

    • Superoxide dismutase (SOD) activity

    • Glutathione (GSH) levels
      Compared to the placebo group (P < 0.05).

  • No significant changes were found in:

    • Catalase (CAT)

    • Glutathione peroxidase (GPx) between the groups.

  • HRW consumption significantly reduced serum malondialdehyde (MDA) levels after the intervention:

    • −25.8% in the HRW group vs. +11.7% in the placebo group (P < 0.001)


Conclusion

Hydrogen-rich water may serve as a novel strategy for preventing oxidative stress or minimizing its harmful effects by enhancing the capacity of antioxidant enzymes in healthy men. However, the exact mechanism of its antioxidant action remains unknown and requires further study.

The Impact of Hydrogen Inhalation Therapy on Blood Reactive Oxygen Species Levels: A Randomized Controlled Study

Chair, M.; Alaani, H.; Fahrioglu, S.L.; Ben Hamda, C.; Fahrioglu, U.; Degheidy, T.
Source: Free Radical Biology and Medicine, Volume 222 (2024), pp. 601–606


Background

Reactive oxygen species (ROS) play a critical role in physiological processes. However, an imbalance favoring ROS over antioxidants leads to oxidative stress, which is associated with a wide range of pathological conditions. Molecular hydrogen (H₂), due to its exceptional permeability and selective antioxidant capacity, has emerged as a promising therapeutic agent. Hydrogen inhalation therapy (HIT) is gaining attention as a novel approach for combating oxidative stress.


Objective

This randomized controlled study aimed to evaluate the efficacy of hydrogen inhalation therapy in reducing blood ROS levels in individuals with elevated oxidative stress.


Methods

  • Participants: 37 individuals with elevated ROS levels (d-ROM value > 350 U.CARR).

  • Study Design: Randomized controlled trial.

  • Groups:

    • Test group: Received hydrogen inhalation therapy (HIT).

    • Control group: Did not receive HIT.

  • Measurements:

    • Blood ROS levels were measured immediately after treatment and 24 hours later for the HIT group.

    • The results were compared with those of the control group.


Results

The test group demonstrated a significant reduction in blood ROS levels post-treatment, both immediately and after 24 hours. The control group showed no significant change.


Conclusion

Hydrogen inhalation therapy (HIT) effectively reduces oxidative stress by lowering blood ROS levels. These findings support HIT as a viable strategy for managing conditions related to oxidative stress.

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Animals

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Effects of Hydrogen-Rich Water on Aging Periodontal Tissues in Rats

Tomofuji, T.; Kawabata, Y.; Kasuyama, K.; Endo, Y.; Yoneda, T.; Yamane, M.; Azuma, T.; Ekuni, D.; Morita, M.
Source: Scientific Reports, Volume 4 (2014)
DOI: https://doi.org/10.1038/srep05534


Background

Oxidative damage contributes to age-related inflammatory responses. The antioxidant effects of hydrogen-rich water (HRW) suppress oxidative damage, which may help inhibit age-related inflammation.


Objective

To investigate the effects of drinking hydrogen-rich water on aging periodontal tissues in healthy rats.


Methodology

  • Subjects: 12 male Fischer 344 rats, 4 months old

  • Groups:

    • Experimental group: treated with hydrogen-rich water (HRW)

    • Control group: treated with distilled water

  • Duration: From age 4 months until 16 months

  • Intervention: Continuous consumption of either HRW or distilled water


Results

  • The experimental group exhibited less oxidative damage in the periodontal tissues at 16 months compared to the control group.

  • No significant difference was observed in the protein expression of interleukin-1β between the groups.

  • However, gene expression of Nod-like receptor protein 3 (NLRP3), an inflammation-related marker, was upregulated in the experimental group compared to controls.


Conclusion

Drinking hydrogen-rich water may exert anti-aging effects by reducing oxidative damage in periodontal tissues. However, it does not significantly impact inflammatory responses in healthy rats.

Hydrogen-Rich Water Attenuates Experimental Periodontitis in a Rat Model

Kasuyama, K.; Tomofuji, T.; Ekuni, D.; Tamaki, N.; Azuma, T.; Irie, K.; Endo, Y.; Morita, M.
Source: Journal of Clinical Periodontology, Volume 38, Issue 12 (2011)
DOI: https://doi.org/10.1111/j.1600-051X.2011.01801.x


Background

Reactive oxygen species (ROS) contribute to the development of periodontitis. Since molecular hydrogen can act as a ROS scavenger, this study investigated the effects of hydrogen-rich water (HRW) on periodontitis in a rat model.


Methodology

  • Induction of periodontitis: Ligatures were placed around the maxillary molars of rats for 4 weeks to induce periodontitis.

  • Treatment: Rats were given drinking water with or without hydrogen-rich water.


Results

  • Rats with periodontitis treated with pure water showed a time-dependent increase in serum ROS levels.

  • Compared to healthy controls, rats with ligature-induced periodontitis and given pure water exhibited:

    • Polymorphonuclear leukocyte infiltration

    • Alveolar bone loss after 4 weeks

  • Hydrogen-rich water treatment:

    • Suppressed the serum ROS increase

    • Reduced expression of oxidative stress markers (8-hydroxydeoxyguanosine and nitrotyrosine) in periodontal tissues

    • Inhibited leukocyte infiltration and osteoclast differentiation during periodontitis progression

    • Downregulated inflammatory signaling pathways, including mitogen-activated protein kinases (MAPKs)


Conclusion

Consumption of hydrogen-rich water may help suppress the progression of periodontitis by reducing oxidative stress in gingival tissues.

Hydrogen-rich water attenuates oxidative stress in rats with traumatic brain injury via Nrf2 pathway

Yuan, J.; Wang, D.F.; Liu, Y.; Chen, X.J.; Zhang, H.L.; Shen, F.; Liu, X.; Fu, J.Q.
Source: Journal of Surgical Research, Volume 228 (2018)
DOI: https://doi.org/10.1016/j.jss.2018.03.043


Background

Recent studies have identified oxidative stress as a key player in the pathogenesis of traumatic brain injury (TBI), making it a potential therapeutic target. Hydrogen-rich water (HRW), due to its antioxidant properties, has demonstrated neuroprotective effects in various neurological disorders. However, the underlying mechanisms of its impact on TBI remain unclear.


Objective

To assess the neuroprotective role of HRW in rats with TBI and to elucidate the potential mechanisms, particularly the involvement of the Nrf2 signaling pathway.


Methodology

  • Model construction: TBI was induced in rats using a modified Feeney weight-drop model.

  • Part 1: Oxidative stress markers (catalase [CAT], glutathione peroxidase [GPx], and malondialdehyde [MDA]) were measured. Nrf2 levels were analyzed to explore its role in HRW’s neuroprotective effect.

  • Part 2: Rats were divided into 3 groups: Sham, TBI, and TBI + HRW.

    • 20 rats per group were monitored for 7-day survival rates.

    • 6 rats per group were assessed for neurological severity score and sacrificed 24h post-injury.

    • GPx, CAT, and MDA were measured using spectrophotometry.

    • Nrf2 and its downstream effectors (HO-1, NQO1) were analyzed via western blotting, RT-PCR, and immunohistochemistry.


Results

  • Oxidative stress markers:

    • GPx and CAT activity significantly decreased.

    • MDA levels peaked 24h post-TBI.

  • Nrf2 expression:

    • Nuclear Nrf2 levels increased in the TBI group, but without significant mRNA changes.

    • HRW administration promoted nuclear translocation of Nrf2 and upregulation of HO-1 and NQO1.

  • Outcomes:

    • HRW improved 7-day survival rates.

    • Reduced neurological deficits.

    • Decreased intracellular oxidative stress.


Conclusion

Hydrogen-rich water provides neuroprotective effects against traumatic brain injury by reducing oxidative stress and activating the Nrf2 pathway, suggesting its therapeutic potential in TBI management.

The protective effect of hydrogen-rich water on rats with type 2 diabetes mellitus

Zheng, M.N.; Yu, H.; Xue, Y.; Yang, T.; Tu, Q.F.; Xiong, K.Q.; Deng, D.H.; Lu, L.; Huang, N.
Source: Molecular and Cellular Biochemistry, Volume 476, Issue 8 (2021)
DOI: https://doi.org/10.1007/s11010-021-04145-x


Background

Hydrogen-rich water (HW) has been reported to exert beneficial effects in patients with diabetes. However, systematic investigation into its mechanisms and efficacy using a suitable animal model was needed.


Objective

To investigate the protective effects of drinking HW on lipid and glucose metabolism, oxidative stress, and inflammation in a rat model of type 2 diabetes mellitus (T2DM).


Methodology

  • T2DM was induced in rats using a high-fat diet combined with low-dose streptozotocin (STZ) injection.

  • Rats were divided into:

    • T2DM + HW group: fed high-fat diet, drank hydrogen-rich water (≥1.0 ppm),

    • T2DM + distilled water group: same diet, drank distilled water,

    • Normal control group: fed standard diet, drank distilled water (n ≥ 10).

  • After 3 weeks of drinking either HW or distilled water, several biomarkers of glucose/lipid metabolism, oxidative stress, and inflammation were measured.

  • Organs including liver, kidneys, and spleen were examined histologically using HE and Oil Red O staining.


Results

  • Drinking HW:

    • Suppressed elevations in blood glucose and total cholesterol.

    • Reduced oxidative stress and systemic inflammation.

    • Alleviated tissue dysfunction in the liver, kidneys, and spleen caused by hyperglycemia.


Conclusion

This study demonstrates that hydrogen-rich water may help improve the metabolic and inflammatory profile in individuals with type 2 diabetes mellitus. Its daily use as drinking water could provide a simple and effective supportive treatment strategy.

Oral Hydrogen-Rich Water Alleviates Oxalate-Induced Kidney Injury by Suppressing Oxidative Stress, Inflammation, and Fibrosis

Si, Y.C.; Liu, L.L.; Cheng, J.; Zhao, T.T.; Zhou, Q.; Yu, J.P.; Chen, W.; Ding, J.R.; Sun, X.J.; Lu, H.T.; Guo, Z.Y.
Source: Frontiers in Medicine, Volume 8 (2021)
DOI: https://doi.org/10.3389/fmed.2021.713536


Objective

To investigate the therapeutic effects and potential mechanisms of hydrogen-rich water (HRW) against oxalate-induced kidney injury.


Methodology

  • A mouse model of calcium oxalate (CaOx) crystallization was induced by administering a soluble oxalate diet.

  • Histological assessments were performed to evaluate crystal deposition, tubular injury, fibrosis, and reactive oxygen species (ROS) production in renal tissues.

  • Serum biomarkers related to kidney injury, inflammation, and oxidative stress were measured using commercial kits.

  • RNA sequencing (RNA-seq) was conducted to identify differentially expressed genes and potential signaling pathways.

  • Western blotting and immunohistochemistry were used to assess the expression of key molecules in the identified pathways.


Results

  • HRW alleviated crystal deposition, tubular damage, fibrosis, and ROS overproduction in the kidneys of oxalate-fed mice.

  • Elevated serum indicators of renal injury, inflammation, and oxidative stress induced by the oxalate diet were reduced by HRW treatment.

  • RNA-seq identified 3,566 differentially expressed genes, and pathway enrichment analysis highlighted PI3K/AKT, NF-κB, and TGF-β signaling as relevant pathways.

  • Expression of pathway-related proteins:

    • PI3K-AKT: PI3K, AKT, and p-AKT

    • NF-κB: NF-κB p65, p-NF-κB p65, NLRP3, and IL-1β

    • TGF-β: TGF-β, TGF-β RI, TGF-β RII, p-Smad2, and p-Smad3
      — were upregulated by the oxalate diet and suppressed by HRW.


Conclusion

Oral administration of hydrogen-rich water may mitigate oxalate-induced kidney damage through its antioxidant, anti-inflammatory, and anti-fibrotic properties, mainly by inhibiting PI3K/AKT, NF-κB, and TGF-β pathways.

Preventive Effects of Drinking Hydrogen-Rich Water on Gingival Oxidative Stress and Alveolar Bone Resorption in Rats Fed a High-Fat Diet

Yoneda, T.; Tomofuji, T.; Kunitomo, M.; Ekuni, D.; Irie, K.; Azuma, T.; Machida, T.; Miyai, H.; Fujimori, K.; Morita, M.
Source: Nutrients, Volume 9, Issue 1 (2017)
DOI: https://doi.org/10.3390/nu9010064


Background

Obesity induces gingival oxidative stress, which contributes to the progression of alveolar bone resorption. The antioxidant effects of hydrogen-rich water (HRW) may help alleviate gingival oxidative stress and prevent bone loss in obese individuals.


Objective

To examine whether drinking hydrogen-rich water can suppress gingival oxidative stress and alveolar bone resorption in obese rats fed a high-fat diet.


Methods

  • Subjects: Male Fischer 344 rats (n = 18)

  • Groups:

    • Control group: Standard diet + distilled water

    • High-fat diet (HFD) + distilled water

    • High-fat diet + hydrogen-rich water

  • Endpoints:

    • Oxidative stress assessed by 8-hydroxydeoxyguanosine (8-OHdG) levels in gingiva

    • Alveolar bone mineral density (BMD) analyzed using micro-computed tomography (micro-CT)


Results

  • Rats fed an HFD showed increased 8-OHdG levels in gingiva and decreased alveolar bone density compared to the control group.

  • Drinking hydrogen-rich water:

    • Suppressed weight gain

    • Reduced gingival oxidative stress (8-OHdG)

    • Attenuated alveolar bone resorption


Conclusion

These findings suggest that hydrogen-rich water may reduce gingival oxidative stress and prevent alveolar bone resorption in obese individuals by limiting obesity-related metabolic disturbances.

Hydrogen-rich water partially alleviate inflammation, oxidative stress and intestinal flora dysbiosis in DSS-induced chronic ulcerative colitis mice

Song, L.H.; Zhang, Y.; Zhu, C.; Ding, X.W.; Yang, L.; Yan, H.L.
Source: Advances in Medical Sciences, Volume 67, Issue 1 (2021)
DOI: https://doi.org/10.1016/j.advms.2021.02.001


Background

Oxidative damage and intestinal dysbiosis play a significant role in the progression of chronic ulcerative colitis (UC). Molecular hydrogen, known for its antioxidant and anti-inflammatory properties, may provide therapeutic benefits.


Objective

To investigate the effects and mechanisms of hydrogen-rich water (HRW) in a chronic UC model induced by dextran sulfate sodium (DSS) in mice.


Methods

  • Subjects: Male C57BL/6 mice (7 weeks old, ~19.6g)

  • Groups:

    • Normal control (NC)

    • DSS-induced colitis group (DSS)

    • DSS + hydrogen-rich water group (DSS + HRW; 0.8 ppm)

  • UC Model Induction:

    • Three DSS treatment cycles

      • First two: 2.5% DSS for 5 days → 16 days water

      • Third: 2% DSS for 4 days → 10 days water

  • HRW was administered daily to the DSS + HRW group throughout the experiment.


Results

  • Clinical signs of colitis developed in DSS-treated mice.

  • HRW treatment:

    • Partially improved clinical symptoms and histopathological changes

    • Increased glutathione (GSH) levels

    • Decreased tumor necrosis factor-alpha (TNF-α) levels

    • Significantly inhibited pathogenic bacteria such as Enterococcus faecalis, Clostridium perfringens, and Bacteroides fragilis (P < 0.05 vs. DSS)

    • Restored microbiota composition toward control levels

  • Microarray analysis:

    • 252 genes significantly altered by HRW

    • 17 inflammation-related genes affected, including 9 interferon-stimulated genes (ISGs)


Conclusion

Hydrogen-rich water can partially mitigate inflammation, oxidative stress, and intestinal dysbiosis in a DSS-induced model of chronic ulcerative colitis, potentially through modulation of inflammatory gene expression and gut microbiota balance.

Hydrogen-rich pure water prevents superoxide formation in brain slices of vitamin C-depleted SMP30/GNL knockout mice

Sato, Y.; Kajiyama, S.; Amano, A.; Kondo, Y.; Sasaki, T.; Handa, S.; Takahashi, R.; Fukui, M.; Hasegawa, G.; Nakamura, N.; Fujinawa, H.; Mori, T.; Ohta, M.; Obayashi, H.; Maruyama, N.; Ishigami, A.
Source: Biochemical and Biophysical Research Communications, Volume 375, Issue 3 (2008)
DOI: https://doi.org/10.1016/j.bbrc.2008.08.003


Background

Hydrogen is a well-established antioxidant known for preventing acute oxidative stress. However, its mechanism of action in the brain remains unclear.


Objective

To clarify the mechanism by which molecular hydrogen (H₂) acts in the brain, especially in the context of oxidative stress and superoxide production.


Methods

  • Animal Model: SMP30/GNL knockout (KO) mice, which cannot synthesize vitamin C (VC)—a known antioxidant.

  • Study Groups:

    • H₂ group: received hydrogen-rich pure water

    • VC group: received vitamin C supplementation

    • H₂O group: received pure water without hydrogen

  • Duration: 33 days of treatment.

  • Vitamin C levels in the H₂ and H₂O groups were less than 6% of those in the VC group, confirming VC depletion.

  • Brain slices from these groups were subjected to hypoxia-reoxygenation treatment, mimicking ischemia-reperfusion injury.

  • Lucigenin-enhanced chemiluminescence imaging was used in real-time to detect superoxide formation in brain slices.


Results

  • The H₂-treated group showed a 27.2% reduction in superoxide formation during ischemia-reperfusion compared to the H₂O group.

  • This reduction occurred despite extremely low vitamin C levels, highlighting the independent antioxidant role of hydrogen.


Conclusion

Hydrogen-rich pure water functions as a powerful antioxidant in brain tissue and effectively prevents superoxide production, even in the absence of vitamin C. These findings support its neuroprotective potential under oxidative stress conditions.

Effects of hydrogen-rich water on aging periodontal tissues in rats

Tomofuji, T.; Kawabata, Y.; Kasuyama, K.; Endo, Y.; Yoneda, T.; Yamane, M.; Azuma, T.; Ekuni, D.; Morita, M.
Source: Scientific Reports, Volume 4 (2014)
DOI: https://doi.org/10.1038/srep05534


Background

Oxidative damage is known to contribute to age-related inflammatory responses. Given that hydrogen-rich water (HRW) has antioxidant properties that suppress oxidative stress, it may help inhibit inflammation associated with aging.


Objective

To evaluate the effects of long-term consumption of hydrogen-rich water on aging periodontal tissues in otherwise healthy rats.


Methods

  • Subjects: Male Fischer 344 rats (4 months old; n = 12)

  • Groups:

    • Experimental group: Received hydrogen-rich water (HRW)

    • Control group: Received distilled water

  • Duration: Both groups were maintained on their respective water regimens until 16 months of age, simulating aging.

  • Evaluations:

    • Oxidative damage in periodontal tissues

    • Protein expression of interleukin-1 beta (IL-1β)

    • Gene expression of Nod-like receptor protein 3 (NLRP3), a marker of inflammation


Results

  • The experimental group showed reduced oxidative damage in periodontal tissues at 16 months of age compared to controls.

  • There was no significant difference in IL-1β protein expression between groups.

  • However, NLRP3 gene expression was increased in the HRW group relative to the control, indicating some activation of inflammation-related genes.


Conclusion

Long-term drinking of hydrogen-rich water appears to exert anti-aging effects by reducing oxidative damage in periodontal tissues. However, it does not significantly alter inflammatory protein responses in healthy aging rats. These findings suggest HRW may be protective against oxidative stress in periodontal aging, but its influence on inflammation in healthy conditions is limited.

Hydrogen-rich water alleviates constipation by attenuating oxidative stress through the sirtuin1/nuclear factor-erythroid-2-related factor 2/heme oxygenase-1 signaling pathway

Chen, K.D.; Wang, K.L.; Chen, C.; Zhu, Y.J.; Tang, W.W.; Wang, Y.J.; Chen, Z.P.; He, L.H.; Chen, Y.G.; Zhang, W.
Source: World Journal of Gastroenterology, Volume 30, Issue 20 (2024)


Background

Constipation is a prevalent functional gastrointestinal disorder that significantly impacts quality of life and imposes a considerable healthcare burden. Oxidative stress plays a central role in colonic dysmotility and underlies the pathophysiology of constipation. Therefore, hydrogen therapy—a novel and promising intervention—has been proposed as a safe and effective treatment for constipation.


Objective

To determine whether hydrogen-rich water (HRW) alleviates constipation and to explore the underlying mechanisms involved.


Methods

  • Animal Model: Constipation was induced in Sprague-Dawley rats using oral loperamide.

  • Treatment: Rats were given free access to HRW.

  • Assessments:

    • 24-hour fecal output

    • Fecal water content

    • Intestinal transit speed (measured with charcoal meal)

  • Microbiota & Metabolomics:

    • 16S rDNA gene sequencing of fecal samples

    • Untargeted serum metabolomics

    • Evaluation of malondialdehyde (MDA) and superoxide dismutase (SOD) levels

  • Histology & Molecular Analysis:

    • Staining (H&E, Alcian blue-PAS) for mucus thickness

    • Immunofluorescence for ROS

    • Immunohistochemistry for c-kit, PGP 9.5, SIRT1, Nrf2, and HO-1

    • Real-time PCR and Western blot for SIRT1/Nrf2/HO-1 expression

  • Mechanistic Validation:

    • Rescue experiment using SIRT1 inhibitor EX527

    • NCM460 cells (human colon cells) exposed to H₂O₂ and treated with key metabolites (beta-leucine and traumatic acid) to evaluate ROS and SIRT1 expression


Results

HRW significantly:

  • Increased fecal volume, water content, and intestinal transit

  • Improved mucus layer thickness, c-kit expression, and enteric neuron count

  • Modulated gut microbiota composition and serum metabolite profiles

  • Reduced intestinal oxidative stress via SIRT1/Nrf2/HO-1 pathway activation

  • These effects were reversed by SIRT1 inhibition

  • In vitro, beta-leucine and traumatic acid mitigated H₂O₂-induced oxidative stress in NCM460 cells by upregulating SIRT1


Conclusion

Hydrogen-rich water alleviates constipation by attenuating intestinal oxidative stress via the SIRT1/Nrf2/HO-1 signaling pathway, while also modulating gut microbiota and key serum metabolites. Beta-leucine and traumatic acid appear to be potential mediators that activate SIRT1 and reduce oxidative damage.

Hydrogen-Rich Water Intake Accelerates Oral Palatal Wound Healing via Activation of the Nrf2/Antioxidant Defense Pathways in a Rat Model

Tamaki, N.; Orihuela-Campos, R.C.; Fukui, M.; Ito, H.O.
Source: Oxidative Medicine and Cellular Longevity, Volume 2016 (2016), Article ID 5679040
https://doi.org/10.1155/2016/5679040


Background

Wound healing is a biological process aimed at restoring tissue integrity and function. Inflammatory cytokines, growth factors, and oxidative stress are crucial components in this process. The transcription factor Nrf2 plays a key role in regulating antioxidant defense mechanisms that counteract oxidative stress during tissue repair.


Objective

This study investigated whether oral intake of hydrogen-rich water (HRW) promotes Nrf2 pathway activation in palatal tissues of rats. The aim was to assess whether this activation reduces systemic oxidative stress and proinflammatory cytokine levels while enhancing the expression of wound healing-related genes.


Methods

  • A circular excisional wound was created in the palatal (roof of the mouth) region of rats.

  • Animals were divided into two groups:

    • One received hydrogen-rich water,

    • The other distilled water (control).

  • The wound healing process and related molecular markers were then monitored.


Results

  • Rats consuming HRW exhibited accelerated wound closure compared to controls.

  • Nrf2 pathway was upregulated in palatal tissues of the HRW group.

  • HRW intake resulted in:

    • Reduced systemic oxidative stress

    • Increased antioxidant activity

    • Decreased levels of proinflammatory cytokines

    • Enhanced expression of healing-associated factors


Conclusion

Oral intake of hydrogen-rich water promoted wound healing in the palatal region of rats by activating the Nrf2 antioxidant defense pathway, reducing oxidative stress, and suppressing inflammatory cytokines. These findings support the hypothesis that hydrogen-rich water has therapeutic potential in enhancing tissue repair and recovery via molecular antioxidant signaling.