
COVID-19 is caused by a viral infection, and can be a lethal disease to the immunocompromised. HydroShot is a popular functional health beverage that is infused with molecular hydrogen, and contains the nitric-oxide-producing amino acid citrulline. Numerous pre-clinical and clinical studies have demonstrated that the gaseous-signaling molecules, molecular hydrogen and nitric oxide, have anti-oxidant, anti-inflammatory, and immuno-modulating benefits. Although nitric oxide can directly kill pathogens, it is critical for immune function, can inhibit viral replication, and can also induce lethal cellular injury under stressed conditions. In contrast, molecular hydrogen has a very high safety profile and helps regulate nitric oxide production, metabolism, and attenuates its harmful effects. Both of these molecules are being used separately in clinical trials for COVID-19 patients. This article briefly reviews the scientific literature on these two molecules, and then presents preliminary data on HydroShot including increased blood flow and wound healing. based on the pre-clinical, clinical, and this preliminary data, HydroShot is recommended for clinical testing/application in addition to its frequent consumer use.
Molecular hydrogen and nitric oxide combination
Nitric oxide, as a reactive free radical, encourages either cell survival or cell death depending on its concentration, location, and timing of production. Importantly, molecular hydrogen regulates nitric oxide (NO•) levels via both reducing excessive production (e.g. suppressing iNOS activity) and also increasing production (e.g. enhancing eNOS activity) . Therefore, NO• regulation is critical to potentiate its beneficial therapeutic effects and mitigate its harmful effects. NO• reacts nearly instantaneously with superoxide (O2•-) to form pernicious peroxynitrite (ONOO-), which is an extremely oxidative and cytotoxic molecule [25]. The reaction with O2•- directly lowers the availability of circulating NO•, which further exacerbates the diseased condition. The radicals, O2•- and NO•, have important beneficial roles in the immune system including killing pathogens, inhibiting their replication, and regulating the inflammatory response. However, under stressful conditions, such as with COVID-19 and its progression to pneumonia, multi-organ failure, and other sequelae, these molecules lose their critical regulation and contribute to the etiology and pathology of these diseases forming both ONOO- and the extremely toxic hydroxyl radical (•OH) . Molecular hydrogen has been demonstrated to favorably regulate the production of both O2•- and NO• by influencing NADPH oxidase, and the NOS enzymes (iNOS, eNOS and nNOS) . Additionally, H2 can effectively reduce the toxic hydroxyl and peroxynitrite oxidants [4]. Furthermore, combination therapy of NO• and H2 demonstrated a synergistic effect in an acute lung-injury model such as what can occur with COVID-19. The treatment significantly attenuated lung neutrophil recruitment, inflammation, and apoptosis induced by LPS and polymicrobial sepsis. Moreover, H2 prevented NO•-induced damage as evidenced by the elimination of nitrotyrosine levels seen with NO• therapy alone. This adds credence to the concept that oral ingestion of HydroShot, a hydrogen-infused functional beverage containing NO•-producing citrulline, may provide significant benefits to the immune system.
HydroShot preliminary data
Increased nitric oxide production: Due to the nitric-oxide stimulating capability of HydroShot, its ingestion accordingly increases endogenous production of nitric oxide as seen in Figure 2. The nitric oxide levels were increased from being depleted at baseline to the optimal level in 30 min, followed by a slight gradual trend upwards at hours 1, 3, 5 and 8 from baseline testing (Figure 2a). Since the test strips are not quantitative and have upper and lower limits, the absolute changes, peak, and duration are unknown.
Similarly, using the FDA approved NIOX VERO machine (Aerocrine AB, Solna, Sweden) the increased nitric oxide levels are also observed (Figure 2b). NIOX VERO is a medical device that measures fractional exhaled nitric oxide (FeNO•), which is a commonly used to help diagnose and manage asthma in children and adults. The baseline FeNO• of 12 subjects (six men and six women; age 45.3 ± 21.1) were obtained, and then they ingested H2Bev H2-infused NO•-stimulating product and their FeNO• levels were measured again 45 min later. The product resulted an average of 202.49% increase in FeNO• levels. Interestingly, when HRW was ingested alone, the NO• levels slightly decreased. This may be due to hydrogen’s ability to influence the regulation of NO• production and decrease its overexpression. Asthmatic patients present with abnormally elevated NO• levels due to an overactive immune response. Molecular hydrogen has been demonstrated to decrease airway inflammation in allergic asthmatic mice. Anecdotally, asthmatic consumers have reported beneficial relief from ingestion of molecular hydrogen, but cite greater therapeutic effects from ingestion of the NO•-stimulating product. For example, a 12-year-old female reported an average Peak Flow Meter Reading at baseline of 220 L/min. However, following 15 min of product ingestion, the flow meter increased by 15%. Finally, after three months of daily use, the baseline flow meter increased to 350 mL/min (160% increase).
Discussion and conclusion
The biomedical effects of molecular hydrogen and nitric oxide are well-substantiated in the scientific literature. Indeed, these molecules have been demonstrated in pre-clinical and clinical studies to exert a wide range of therapeutic effects. In line with our data, previous research has demonstrated that high-concentration HRW could improve endothelial function and significantly increase flow-mediated dilation of the brachial artery. Similarly, the increased healing effects of HydroShot are corroborated by previous research where administration of HRW via tube-feeding to severely hospitalized elderly patients with pressure ulcers resulted in wound size reduction, and a faster recovery. However, when compared to the preliminary data from HydroShot, these results suggest that the addition of citrulline significantly increases the therapeutic action (i.e. blood flow and wound healing) of the beverage.
At the cellular level, COVID-19 and similar diseases are characterized by dysregulated inflammatory response of the immune system, excessive oxidative stress, and impaired cellular function. Nitric oxide plays a dual role in actively improving the immune system, destroying pathogens, inhibiting their replication, and also in promoting cellular oxidative damage and promotion of excessive inflammation. However, since nitric oxide plays a key role in healthy cellular function, inhibiting or activating the NOS enzymes via pharmacological intervention similarly has both beneficial and harmful consequences. In contrast, molecular hydrogen is a novel and safe medical gas that affords therapeutic effects by regulating redox status and the inflammatory response. In addition to the clinically relevant antioxidant and anti-inflammatory effects of H2, it also has the ability to regulate nitric oxide production and attenuate its adverse effects.
Currently, inhalation of H2 gas is being used in at least four clinical studies [RefA]. Dr. Zhong Nanshan, the epidemiologist who discovered the SARS coronavirus in 2003, recently recommended the inhalation of molecular hydrogen for COVID-19 patients based on the preliminary clinical data [RefC]. Hydrogen treatment resulted in significant amelioration of dyspnea in most patients with COVID-19 in the pilot investigation. Correspondingly, hydrogen inhalation is included as a recommendation by the National Health Commission of The People’s Republic of China’s protocol for COVID-19 [RefD]. Although their research is on inhalation of molecular hydrogen, ingestion of HRW is also effective, and, at least in some cases, more effective. Moreover, the addition of citrulline to boost nitric oxide levels, both of which can be in deficiency during late stages of COVID-19, further give credence to its observed therapeutic effects in the presented preliminary data. In conclusion, due to the urgency of the COVID-19 pandemic, and the simplicity, safety, and clinical viability of HydroShot, it is recommended that HydroShot, in addition to its frequent consumer use as a healthy beverage, be clinically investigated for its potential use in treating COVID-19 patients
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