1. Introduction
Obesity, type 2 diabetes mellitus (T2DM), and cardiovascular diseases (CVDs) are
leading causes of global mortality [1, 2, 3]. These conditions share common risk
factors including, an unhealthy diet, tobacco and alcohol use, and insufficient
physical activity [1]. The impact of a western diet is notable, inducing
oxidative stress compromising the body’s antioxidant capacity and instigating
inflammation [4, 5]. This systemic inflammation can specifically affect the beta
cells of the pancreas, hepatocyte low-density lipoprotein (LDL) receptors,
endothelium, neurons, and osteocytes [6, 7]. These factors collectively raise the
susceptibility to CVDs and T2DM, ultimately resulting in heightened mortality
[6, 7].
The role of antioxidants in mitigating oxidative stress associated with these
diseases remains an area of ongoing research. One hypothesis is that a healthy
diet can balance oxidative stress levels, maintain cell and tissue homeostasis,
and consequently reduce inflammation leading to a decreased risk of CVDs and
metabolic disorders [8, 9]. Western diets (WD) exacerbate oxidative stress by
elevating the levels of protein carbonylation and lipid peroxidation [4, 5] while
decreasing the gut’s production of molecular hydrogen (H), a potential
antioxidant [10, 11]. Conversely, Mediterranean diets (MD) which are rich in
dietary fiber, flavonoids, and omega-3 fatty acids [12], may bolster antioxidant
defenses by facilitating the production of protective molecules like H[ 10, 11, 13].
While the protective mechanisms of H remain unclear, evidence suggests
that H supplementation can reduce oxidative stress and inflammation,
offering protection from CVDs and metabolic diseases [10, 11, 12, 13, 14, 15]. There are several
methods for increasing H including inhaling H gas, drinking
hydrogen-dissolved water (H-water), injecting hydrogen-dissolved saline
(H-saline), taking hydrogen baths, and applying H-saline to the eyes.
This communication aims to highlight the role of H, in the management of
cardio-metabolic diseases (CMDs).
2. Free Radical Stress and Antioxidants in the Pathogenesis of Chronic
Diseases
The combination of WD and environmental factors including pollution, tobacco
smoke, pesticides and pollutants contribute to the generation of free radicals
[4, 5, 10, 11, 14]. In the body inhaled oxygen (O) undergoes single electron
reduction to form superoxide radicals (O) [15]. These radicals can
either propagate further oxidative reactions or transform into other reactive
species such as hydrogen peroxide (HO) and hydroxyl radicals
(•OH) [15]. Free radicals have an unpaired electron, and are
consequently very reactive, requiring a single electron to form a stable electron
shell [15]. These free oxygen radicals scavenge body tissues, leading to
cellular and molecular damage [15]. This activity impacts cells, proteins,
lipoproteins, and DNA, serving as a catalyst for various diseases [15].
The body naturally produces a range of free radicals, reactive oxygen species
(ROS) and reactive nitrogen species from endogenous metabolic processes oxidants,
exposure to environmental toxicities, and disease processes [15]. Maintaining a
balance between free radicals and the body’s antioxidant defenses is critical for
metabolic health, imbalances can elevate oxidative stress, causing tissue damage,
and increasing the risk of conditions including CVDs and T2DM [8, 9, 10, 11, 14].
Interestingly, physiological levels of free radicals can have protective effects
on cells, emphasizing the importance of endogenous antioxidants in neutralizing
free radical-induced tissue damage [4, 5]. Notable endogenous sources of these
toxins include xanthine oxidase, nicotinamide adenine dinucleotide phosphate
(NADPH) oxidase, and electron leakage of electrons from the mitochondrial
respiratory chain, which generate harmful superoxide radicals [16, 17].
3. Diet, Western Diet, Microbiome, and Molecular H
The WD, characterized by a high intake of saturated fats and sugar with a
simultaneous low fiber intake, plays a significant role in the rise of chronic
diseases and mortality rates [18]. Commonly consumed industrially manufactured
ultra processed foods, including carbonated soft drinks, fast foods, industrially
produced breads, or hot dogs have reduced nutritional values [19]. These dietary
habits contribute to elevated cardiovascular risk factors (e.g., dyslipidemia,
hypertension), and obesity or metabolic syndrome (MS) leading to increased
mortality rate [18, 19, 20, 21]. This dietary pattern is increasingly becoming a health
concern, driving a surge in metabolic diseases like diabetes and obesity,
particularly in countries adopting a Western lifestyle [22].
Recent studies highlight the role of gut microbiota in the development and
progression of inflammation, often called metaflammation which is linked to the
development of non-communicable diseases involving immune system dysregulation
[22]. Diets can affect the gut microbiota resulting in alterations to the host’s
physiological responses. Consuming a WD can disrupt the gut’s microbial balance,
leading to dysbiosis and further exacerbating gut inflammation [23].
In contrast, the MD is known for its anti-inflammatory properties, primarily due
to its emphasis on a plant-based, prebiotic-rich foods, such as asparagus,
garlic, onion, leeks, and omega-3 fatty acids [24]. These dietary components
provide nutrients that interact with gut microbiota, and the immune system to
maintain homeostasis [24]. Polyunsaturated fatty acids, fiber, and polyphenols
may reduce the risk of chronic diseases by regulating oxidative stress [24]. The
precursors of these diphenols are found in fiber-rich unrefined grain products,
seeds, beans, peas, and berries [24]. While dietary antioxidants may provide
protection from oxidative damage by neutralizing ROS, translating this into
clinical practice has proven challenging. One issue is that antioxidants
indiscriminately reduce all ROS levels, including those involved in beneficial
physiological signaling [24].
4. Diet as Oxidant and Antioxidant Agent
The combination of a WD with low dietary antioxidant intake leads to an
antioxidant deficiency, along with an increase in free radical stress-induced
tissue damage throughout the body [4, 5, 10, 11, 14]. Noteworthy endogenous
antioxidants such as catalase, superoxide dismutase, and ceruloplasmin are
protective against damage to cholesterol receptors in the hepatocytes, beta cells
of the pancreas, and endothelial damage by inhibiting free radical generation
[8]. There is evidence that the WD is deficient in antioxidant nutrients such as
flavonoids, fiber, and omega-3 fatty acids, leading gut microbes to produce fewer
protective molecules including short chain fatty acids, glucagon like peptides,
and H, which are potential anti-inflammatory agents [4, 5, 11, 14, 15].
Conversely, the MD is rich in antioxidants such as vitamins, minerals,
flavonoids, omega-3 fatty acids, and fiber, can inhibit oxidative stress and
inflammation, thereby reducing the risk of CVDs and T2DM [4, 5, 8, 9]. Antioxidant
rich diets also promoted H production in the gut, which may regulate
circadian variations in blood pressure [8, 11, 14, 15]. In clinical settings, H
has been demonstrated to inhibit free radical stress in subjects with endothelial
dysfunction, CVDs, and T2DM diabetes, that occur, due to oxidative stress [11, 15]
(Fig. 1).
Fig. 1.
The impact of diet on oxidative stress and inflammation: role of
molecular hydrogen in of cardiovascular and metabolic disease development. H, hydrogen.
5. Production of Molecular Hydrogen in the Gut
The gut microbiota plays a crucial role in mitigating the risk of T2DM and CVDs
[25]. Many complex carbohydrates and plant polysaccharides escape digestion in
the gut due to the absence of enzymes [25]. However appropriate microbes can
metabolize these polysaccharides into beneficial short-chain fatty acids (SCFAs)
with potential anti-inflammatory effects such as propionate, butyrate, and
acetate, along with gases like methane and H [25, 26]. H is generated
through fermentation of carbohydrates – such as lactose, lactulose, and fructose
by intestinal bacteria [25, 26]. The primary bacteria involved in H
production are groups such as Bacteroides fragilis, Clostridium
perfringens, and Pseudomonas, all of which are normally present in the
large intestine and possess hydrogenases [25, 26]. This fermentation producing
SCFA typically occurs in the colon [25, 26]. Their concentration tends to be higher
in the proximal colon and lower in the distal colon, despite the latter having a
greater microbial density and elevated gas levels [25, 26]. Besides their local
effects, SCFAs like acetate can influence neural function, offering a potential
pathway for gut-brain interactions [25, 26] (Fig. 2).
Fig. 2.
Mechanism of production and inhibition of molecular hydrogen due
to diets via microbiota in the gut, and its effects on anti-inflammatory
molecules and cardio-metabolic diseases. H, hydrogen; IL4, IL6, and IL10,
interleukin 4, 6, and 10; TGF, transforming growth factor beta; GLP,
glucagon-like peptide; BDNF, brain-derived neurotrophic factor; SCFA, short-chain
fatty acids; NO, nitric oxide; RAAS, renin-angiotensin-aldosterone system; Ang
II, angiotensin II; CAD, coronary artery disease; SMC, smooth muscle cell.
H production in the human gastrointestinal (GI) tract is primarily
dependent on the fermentation of ingestible fibrous substrates by a rich
intestinal flora [27], predominantly located in the colon [24]. The amount of
endogenous H produced through this mechanism generally surpasses that
obtained from consuming H-rich water (HRW). Excess H can be removed
through multiple microbial pathways [11]. In addition to methanogenesis, another
mechanism involves sulphate-reducing bacteria using excess H to convert
sulphate to sulfite. The “keystone pathogen” hypothesis offers an explanation
for the role of specific microbes in disease states, that certain low-abundance
microbial pathogens can disrupt a normally benign microbiota, converting it into
a disease associated, or dysbiotic state [28]. These pivotal microbes, termed
“keystone pathogens”, play a role in creating an environment conducive to
disease, particularly by fostering inflammation [29]. Recent studies substantiate
the idea that these pathogens instigate disease by altering the gut microbiota
[29].
When combined with the proper diet, gut microbiota can generate between 3-9
liters of H in the colon [30]. H is formed as an end product of
polymeric carbohydrate fermentation caried out by members of the
Firmicutes and Bacteroidetes microbial taxa [30]. There are two
primary pathways for H disposal, methanogenesis, and homoacetogenesis, with
the latter being more predominant. H is produced by many members of the gut
microbiota and may be subsequently utilized by cross-feeding microbes for growth
and in the production of larger molecules [31]. H can serve as a substrate
for hydrogenotrophic microbes, which fall into three categories: sulfate-reducing
bacteria, methanogenic archaea, and acetogenic bacteria, which can convert
H into hydrogen sulfide, methane, and acetate, respectively [30, 31, 32]. It is
becoming increasingly clear that H plays a crucial role in GI microbial
metabolism, impacting human nutrition, health, and wellbeing, with a growing body
of evidence suggesting a strong correlation between the volume of Hproduction by intestinal bacteria and various diseases [13].
A pilot study reported that consumption of H-producing milk four hours
prior to exercise significantly decreased creatine kinase and
8-hydroxy-2-deoxyguanosine levels while improving muscle recovery following
exercise [33]. Previous research indicated that acetate facilitates a
microbiome–brain–-cell axis that exacerbates MS [26], while increased
production of metabolites including short chain fatty acids, brain-derived
neurotrophic factor (BDNF), and H enhance metabolism via gut-brain
interaction neural circuits [25]. A healthy gut microbiota is promoted when the
fiber rich MD includes probiotics, increasing H production to levels
measurable in liters [27, 28, 29]. The medical community is increasingly exploring
natural, non-toxic compounds like H for their potential antioxidative roles
in preventing cardiovascular diseases and other chronic conditions [10, 11, 14, 15]. The
evolving understanding of the biological importance of intestinal H has
shifted the perception of its significance. No longer just a byproduct, H
is now viewed as a critical factor in global organ function and homeostasis
[11, 15, 16, 17].
Over the past two decades H has emerged as a versatile antioxidant with
applications across a spectrum of physiological and pathophysiological
conditions. Whether endogenously produced through healthy foods or exogenous
administration via inhalation or HRW, H has shown promise as a potential
antioxidant in a wide range of physiological and pathophysiological processes
[16, 17]. H can inhibit hydroxyl and nitrosyl radicals in the cells and
tissues, causing a marked decline in oxidative stress, leading to a decline in
the inflammation that is marker in the pathogenesis of diabetes and CVDs [17].
Interestingly, Slezák et al. [10] and other researchers [11, 14, 15]
have demonstrated that H can rapidly diffuse into tissues and cells without
disrupting metabolic redox reactions or signaling reactive species (Fig. 3, Ref.
[11]).
Fig. 3.
Mechanism of action of molecular hydrogen in the pathogenesis
and control of cardiovascular and metabolic diseases (Modified from reference
[11], Ichihara et al., Med Gas Res 2015). •OH, hydroxyl radical; ONOO,
peroxinitrite; H, hydrogen; NO, nitric oxide; IL-10, interleukin 10.
In addition to regulating gene expression, H engages in epigenetic
modulation, offering alternative pathways for mitigating oxidative stress-induced
genetic damage, thereby enhancing its anti-inflammatory and anti-apoptotic
capabilities [16, 17]. H also alleviates blood-brain barrier impairment and
improves cognitive dysfunction [34]. Hydrogen therapy has been found to
ameliorate cardiac remodeling [35], dyslipidemia and MS [36] oxygen saturation in
chronic lung disease [37], and in nonsteroidal anti-inflammatory drug
(NSAID)-induced enteropathy [38].
6. Oxidative Stress, Inflammation, Immunomodulation, and Effects of
H
Oxidative stress arises from an the imbalance between the production of reactive
oxygen and nitrogen species and the body’s ability to eliminate reactive
intermediates. Many antioxidants acting through different mechanisms have been
successfully used as a form of therapy, preventing cell damage [10, 39, 40]. While
oxidative stress is a natural part of aging, over 2000 scientific papers
implicate chronic oxidative stress to the development of a whole range of chronic
pathological conditions [39]. Critical macromolecules including DNA, proteins,
and membranes can be damaged by highly reactive hydroxyl and nitrosyl radicals
during periods of oxidative stress [39, 41, 42].
•OH radicals are highly reactive and can interact with virtually any biological
molecule in the vicinity [41]. The scavenging of free radicals can serve both
preventive and therapeutic roles [39]. H, due to its selective reactivity,
stands out as a unique scavenger, reacting only with •OH and
peroxynitrite (ONOO) [39]. Other ROS like superoxide
(O), HO and nitric oxide
(•NO)—which also serve as signaling molecules—remain unaffected
[39]. Furthermore, H indirectly regulates hormones and cytokines through
various signal transduction pathways [39]. During the inflammatory response
immune cells break the homeostatic balance; H inhibits pro-inflammatory
signaling and activates anti-inflammatory signaling [43].
An essential attribute of H is its permeability, enabling rapid
penetration of the cell membrane and dispersion throughout the cytoplasm,
nucleus, and other organelles to confer protective effects [44]. In contrast to
most antioxidant compounds, H can pass through the blood–brain barrier,
and thus far, there have been no reports of cytotoxicity [44]. H also has
no direct effect on body temperature, blood pressure, pH, or pO [44].
H exerts anti-inflammatory and antioxidative effects by directly
interacting with the mitochondrial electron transport chain and neutralizing
oxidative stress [45]. Overall, this alleviates mitochondrial damage, balances
intracellular environmental homeostasis, and protects the transcription of key
regulatory proteins of inflammation [45].
7. Molecular Hydrogen Therapy for Cardiovascular Diseases
Damage to cardiomyocytes, vasculature—including endothelium and smooth muscle
cells—are all CVD risk factors that result in cardiovascular dysfunction [35].
Increases in fibrosis and apoptosis are closely related heart failure [35].
Therefore, novel therapeutic approaches for the treatment of cardiac remodeling
and fibrosis of myocardium are needed to improve the survival rates of patients
with cardiac ischemia. In a rat model of myocardial infarction, H treatment
(inhalation of 2% H for 28 days daily for 3 hours) significantly improved
cardiac function while decreasing the area of fibrosis [35]. Complementary
in vitro experiments also revealed that H therapy mitigated
hypoxia-induced damage to cardiac cells and inhibited the angiotensin II-induced
migration and activation of cardiac fibroblasts [35].
ROS play a significant role in vascular disease development while also
modulating blood vessel vasomotor function [14]. These free radicals neutralize
•NO, converting it into the more harmful peroxynitrite radical [43].
NADPH oxidase (NOX) family proteins, the oxidases that produce HO and superoxide, are
the main source of vascular free radicals [17, 43, 46, 47]. Variations in blood
pressure and flow can impact endothelial function, which is crucial for
maintaining vasomotor tone, as the arterial endothelium actively modulate shear
stress [14]. Accumulated oxidative stress and inflammation can lead to
endothelial dysfunction, predisposing individuals to atherosclerosis and CVDs
[14]. Endothelium-derived relaxing factors (EDRF), such as •NO,
endothelium-derived hyperpolarization factor (EDHF) and prostacyclin are known to
play a crucial role in the development of diet induced vascular dysfunction
[4, 5]. The shear stress activates the NOX proteins—specifically NOX 1, NOX2 and
NOX3 —which are key factors in vascular function [46, 47]. Superoxide radicals,
primarily generated by NOX1 and NOX2 through single electron transfer to H,
rapidly neutralize excess •NO within cells, leading to the production
of peroxynitrite [47]. This compound adversely affects vasodilation mediated by
nitric oxide [47].
In the presence of peroxynitrite, an indication of oxidative dysfunction, there
may be a suppression of endothelial nitric oxide synthase (eNOS) enzyme activity,
leading to reduced NO production [47]. The eNOS oxidation inducing cofactor,
tetrahydrobiopterin (BH4), may be converted to the inactive form
7,8-dihydropterin (BH2). This conversion leads to the uncoupling of eNOS, a
mechanism that generates superoxide radicals [47].
The redox imbalance between •NO and superoxide radical production in
endothelial cells may lead to endothelial dysfunction [14]. Another ROS,
HO have both detrimental and beneficial effects on vascular function.
Although the role of the hydroxyl radical—a byproduct of hydrogen peroxide
decay—is not fully understood, it is known to impair endothelial function. This
impairment can be counteracted by H [14]. A clinical study demonstrated
that H therapy significantly improves flow-mediated dilatation (FMD) in
healthy volunteers suggesting protective effects on vascular function [14]. In
the group receiving high levels of H, FMD increased from 6.80% 1.96% to 7.64% 1.68% (mean SD) compared to a decrease from
8.07% 2.41% to 6.87% 2.94% in the placebo group [14]. These
findings indicate that H may protect vascular tissues from damage induced
by shear stress and hydroxyl radicals, while preserving the beneficial effects of
nitric oxide on vasomotor function. Given that oxidative stress can exacerbate
systemic inflammation and thereby impair the function of cardiomyocytes, beta
cells, and neurons, as well as endothelium, the potential protective roles of
H warrant further investigation [48, 49, 50, 51, 52].
ROS are generated as essential co-factors during oxidative phosphorylation via
electron transfer, a process that occurs in aerobic metabolism [11, 14, 15].
Rheumatoid arthritis (RA) is known to elevate the risk of coronary artery disease
(CAD) and atherosclerosis, which in turn increases mortality from CVD [53]. This
link can be attributed to overlapping inflammatory pathways in both RA and CAD
[53]. Specifically, free radicals and pro-inflammatory cytokines appear to be key
drivers connecting these diseases [54]. These inflammatory mechanisms impact both
the vascular endothelium and joint tissues in arthritis. Endothelial and smooth
muscle cells produce superoxide radicals through NADPH oxidases, including NOX1,
NOX2, NOX4, and NOX5, which are crucial to endothelial function and progression
of atherosclerosis [53, 54, 55]. The oxidation of low-density lipoprotein cholesterol (LDL-C), observed as an intersection between these mechanisms, predisposing
plaque development in atherosclerosis, consequently leading to high CVD risk
[56, 57]. The development of CAD or stroke in patients with arthritis may lead
individuals to be predisposed due to changes in endothelial phenotypic response
to a high quantity of harmful stimuli. Oxidative stress upregulates the
expression of adhesion molecules such as vascular cell adhesion molecule-1
(VCAM-1), intercellular adhesion molecules 1 (ICAM-1), and E-selectin. The
pro-inflammatory cytokines tumor necrosis factor alpha (TNF-),
interleukin 1 (IL-1), interferon-, are also activated in the
pathogenesis of atherosclerosis [53, 54, 55, 56, 57]. Interestingly, vascular dysfunction may
also occur due to up-regulation of TNF- expression alone, leading to
atherosclerosis [58]. In patients with rheumatoid arthritis, anti-TNF-
therapies could reduce the progression of atherosclerosis, indicating that the
pathogenesis of atherosclerosis involves shared TNF/ROS inflammatory
pathways at the crossing between Loop 1 and 2 [58]. Further studies by Slezak and
his group [59, 60, 61, 62] have illustrated the role of H in hypoxic
post-conditioning, radiation-induced heart injury, or acute cardiac injury (Fig. 4, Ref. [42]).
Fig. 4.
Mechanisms of the effects of hydrogen therapy on
cardio-metabolic diseases (Modified from LeBaron et al., 2019, reference
[42]). SMCs, smooth muscle cells; CVDs, cardiovascular diseases; NF-B,
nuclear factor kappa B; TNF-, tumor necrosis factor alpha; ICAM-1,
inter cellular adhesion molecule-1; (INF)-, interferon gamma; IL-1,
interleukin 1 beta; HMG-1, high mobility group box 1 protein; MMP2, matrix
metalloproteinase 2; Bcl-2, B-cell lymphoma 2; Bcl-xL, B-cell lymphoma-extra
large; ASK1, apoptosis signal-regulating kinase 1; MAPK, mitogen-activated
protein kinase; JNK, c-Jun N-terminal kinases; Nrf2, nuclear factor erythroid
2-related factor 2; •OH, hydroxyl radical; ONOO,
peroxinitrite; SOD, superoxide
dismutase; CAT, catalase; GSH, glutathione; H, hydrogen.
8. Effects of Hydrogen in Stroke
The medicinal value of H has been shown by inhalation of 2% H which
can significantly decrease the damage caused during cerebral
ischemia/reperfusion, which in turn are caused by oxidative stress via selective
elimination of •OH and ONOOˉ [63, 64, 65, 66, 67, 68, 69, 70]. Numerous experimental and
clinical studies involving H indicate that therapy produces
anti-oxygenation, anti-inflammation, and anti-apoptosis effects. Since brain
tissue is highly susceptible to cell damage, produced by free radicals and other
markers, H therapy benefits may be easier to demonstrate in patients
predisposed to stroke [63, 64, 65, 66, 67, 68, 69, 70]. A single comprehensive review accounting for the
blood-brain barrier, penetrability, possible side effects, and the molecular
properties of H, should contribute to advancing both clinical and
experimental research and therapies. In clinical studies, upon ischemic stroke
onset, 8.5–30% of patients suffer a hemorrhagic stroke, and the rest experience
an ischemic stroke [64]. In animal studies, small doses of H have been
shown to significantly reduce mortality in cases of brain wide ischemic strokes
[59]. When H was administered to groups with high sugar levels and
transient middle cerebral artery occlusion (tMCAO), it effectively lowered the
risk of brain hemorrhage [59]. Sustained inhalation of 2.9% H for 2 hours
led to a significant reduction in oxidation and nitration byproducts, as well as
in matrix metalloproteinase-9 (MMP-9), suggesting that the blood-brain barrier
was better preserved [59]. Chen et al. [66] proposed that this effect
contributed to the lower occurrence of hemorrhage accompanying cerebral
infarction. In a separate study, mice were subjected to global cerebral
ischemia/reperfusion (I/R) through a 45-minute occlusion of both common carotid
arteries (BCCAO) [64]. Inhalation of 1.3% H-rich air improved the 7-day
survival rate, significantly mitigating neuronal damage, autophagy in the
hippocampal CA1 region, and brain edema. Additionally, the administration of
H led to lower levels of oxidative stress markers 8-hydroxy-2-deoxyguanosine (8-OHdG) and
malondialdehyde in brain tissues [64].
A hemorrhagic stroke is defined as a cerebral hemorrhage followed by compression
and necrosis of brain tissue [67]. Hemorrhagic strokes are typically more
dangerous than ischemic strokes because they involve microglia and inflammatory
cells, which are activated upon hemorrhage, producing free radicals [68]. In a
mouse model of intracerebral hemorrhage, inhalation of 2% H for one hour
significantly reduced the degree of cerebral edema and significantly improved
neural function [69]. Interestingly, these improvements were limited to 72 hours,
suggesting that H inhalation provides protection only in the acute phase of
cerebral hemorrhage [69]. The delay in the peak of neutrophil infiltration and
microglial activation, occurring after 72 hours, might explain why the
anti-oxygenation effects of H were not sufficiently persistent during that
period [69]. Additionally, H’s protective effect on the blood-brain barrier
and its ability to reduce cerebral edema may be attributed to its moderating
influence on mastocyte activity, which is crucial in the initial inflammatory
responses following a stroke [69, 70]. In a study involving rats with acute
hyperglycemia, treatment with H-rich saline was associated with increased
hemorrhagic transformation in a focal ischemia [66]. Meanwhile, a rabbit model of
subarachnoid hemorrhage on brain stem infarction showed the combination of
H and edaravone treatment led to a more significant reduction in recovery
time compared to using edaravone alone [70].
9. Effects of Molecular Hydrogen on Blood Lipoproteins
Increased concentrations of blood lipids and pro-inflammatory cytokines are risk
factors of CVDs. Clinical and experimental studies indicate that H
administration has beneficial lipid-lowering effects. In a case study of 20
patients with MS, HRW (0.9–1.0 L/day) was administered to determine its effects
on biological activities of serum lipoproteins [28, 71]. Following 10 weeks of
treatment, HRW produced a decline in total-cholesterol (TC) and LDL-C concentrations [28, 71]. This was accompanied by a significant decline in
apolipoprotein (apo)E, apoB100, and an improvement in function of high-density
lipoprotein (HDL) [28, 71]. The intake of HRW was associated with rise in
superoxide dismutase (SOD) and a reduction in thiobarbituric acid-reactive
substances (TBARS) in the LDL and serum, important markers of MS [28, 71]. In a
clinical trial, 68 patients with high cholesterol were randomized to either HRW
(0.9 L/day, n = 34) or placebo (n = 34) for a total period of 10 weeks [29, 72].
In the group treated with HRW, the isolated HDL cholesterol demonstrated enhanced
efficacy in promoting adenosine triphosphate (ATP)-dependent cholesterol efflux, specifically related to
the cassette transporter A [29, 72]. Concurrently, there was an increase in plasma
levels of pre--HDL, while HDL-cholesterol concentrations remained
unchanged [29, 72]. Moreover, HRW treatment was associated with improvement in
other HDL functions related to LDL oxidation, specifically inhibition of
pro-inflammatory oxidized-LDL and the protection of endothelial cells from
apoptosis. In addition, therapy with HRW was associated with the improved
down-regulation of total cholesterol (47.06% vs. 17.65%) and LDL-C
(47.06% vs. 23.53%). There was a significant decline in apoB100 with rise in
apoM in the H group. Treatment with H was associated with a marked
decline in the concentrations of multiple pro-inflammatory markers as well as
indicators of oxidative stress in the plasma and HDL particles. The present
results emphasize the potential efficacy of H therapy in the reduction of
cholesterol as well as atherosclerosis.
10. Effects of Molecular Hydrogen in Diabetes Mellitus and Metabolic
Syndrome
MS is characterized by the presence of at least three of the following risk
factors including obesity, diabetes, hypertension, hyperlipidemia, and low HDL
[71]. Free radicals, with or without inflammation, are thought to play key roles
in the development of MS and T2DM [71, 72, 73]. Therapy with HRW has shown promise in
improving glucose and lipid metabolism in individuals with T2DM or glucose
intolerance, conditions which are both linked to oxidative stress [73, 74]. The
effectiveness of HRW (1.5–2 L/day) was examined in an open label, 8-week study
in 20 subjects with potential MS [73]. HRW was generated by inserting a metallic
magnesium stick into drinking water, leading to an H concentration between
0.55–0.65 mM) produced from the following chemical reaction [73]:
Mg + 2HO Mg (OH) + H
The consumption of HRW for 8 weeks resulted in a 39% increase (p
0.05) in the antioxidant enzyme SOD and a 43% decrease (p 0.05) in
TBARS in urine [73]. Furthermore, subjects showed an 8% increase in
HDL-cholesterol and a 13% decrease in total cholesterol/HDL-cholesterol from
baseline to week 4 [73]. There was no change in fasting glucose concentrations
during the 8-week study [73]. Drinking HRW may represent a potentially novel
therapeutic and preventive strategy for MS.
Singh and colleagues [36] conducted a randomized, placebo-controlled trial where
patients with MS with were treated with HRW, showing favorable effects on
multiple parameters following 24 weeks when compared to placebo group (p 0.05, p = 0.309). The results were accompanied by significant
declines in body mass index (BMI) and waist-to-hip ratio (WHR, p
0.05) [36]. In addition, treatment with HRW caused a significant decline in blood
lipids as shown in Table 1 (Ref. [36]).
Table 1.Effects of hydrogen rich water on blood lipoproteins in
patients with metabolic syndrome [36].
Data |
Hydrogen rich water (n = 30) |
Placebo (n = 30) |
Data, mg/dL |
Baseline |
After 24 weeks |
Baseline |
After 24 weeks |
Cholesterol |
187.7 32.4 |
169.2 26.1*** |
184.3 37.4 |
184.4 38.6 |
LDL cholesterol |
109.0 34.4 |
102.5 28.0 |
105.5 42.0 |
106.0 43.3 |
HDL cholesterol |
41.7 4.2 |
40.4 1.8 |
41.8 2.3 |
42.3 2.4 |
VLDL cholesterol |
37.3 17.9 |
28.0 11.3** |
36.8 20.6 |
37.3 20.5 |
Triglycerides |
189.8 93.3 |
142.4 65.0** |
184.4 102.8 |
185.6 101.3 |
C-reactive proteins |
0.5 0.2 |
0.5 0.1* |
0.6 0.5 |
0.6 0.5 |
*** = p value 0.0001, ** = p value 0.01, * = p
value 0.05, by comparison of baseline and after following up using analysis
of variance (Modified from reference [36]). LDL, low density lipoprotein; HDL,
high density lipoprotein; VLDL, very-low-density lipoprotein.
Treatment with HRW also reduced fasting blood glucose after 24 weeks, along with
a significant decline in glycated haemoglobin (HbA1C) (12%, p 0.05) compared to baseline
levels and placebo group [36]. Treatment with HRW also reduced the markers of
inflammation: TNF-, and IL-6 (p 0.05) [36]. While
oxidation markers showed a significant decline, there were increases in vitamins
C and E in the H group [36]. Serum levels of angiotensin converting enzyme
were significantly decreased whereas serum nitrite level showed significant
increases (Table 2, Ref. [36]), which may lead to declines in blood pressure.
Table 2.Effect of hydrogen rich water on glycaemia, oxidative stress,
and cytokines in patients with metabolic syndrome [36].
|
Hydrogen rich water (n = 30) |
Placebo (n = 30) |
Data, mg/dL |
Baseline |
After 24 weeks |
Baseline |
After 24 weeks |
Fasting blood glucose |
121.5 61.0 |
103.1 33.0* |
123.9 43.4 |
126.4 42.3 |
HbA1c, % |
5.8 0.9 |
5.1 0.2*** |
6.2 1.2 |
6.1 1.2 |
TNF- |
4.8 1.2 |
3.9 0.6*** |
4.8 1.3 |
4.8 1.3 |
IL-6 |
1.9 0.7 |
1.6 0.2** |
1.6 0.6 |
1.7 0.6 |
TBARS |
2.5 0.3 |
1.6 0.3* |
2.5 0.3 |
2.5 0.3 |
Malondialdehyde |
3.4 0.2 |
2.7 0.2*** |
3.4 0.2 |
3.5 0.2 |
Diene conjugates |
27.8 1.0 |
26.7 0.5*** |
28.3 0.8 |
28.3 0.8 |
Vitamin E |
23.0 2.3 |
26.8 1.9*** |
23.0 1.5 |
23.1 1.1 |
Vitamin C |
20.7 2.5 |
24.2 1.8*** |
20.7 2.5 |
20.8 2.4 |
Nitrite |
0.63 0.06 |
0.68 0.06*** |
0.66 0.04 |
0.65 0.03 |
Angiotensin converting enzyme |
85.2 7.8 |
80.7 5.8*** |
84.5 8.8 |
83.8 8.7 |
*** = p value 0.0001, ** = p value 0.01, * = p
value 0.05, by comparison of baseline and after follow up using analysis of
variance (Modified from reference [36]). HbA1c, glycated hemoglobin;
TNF-, tumor necrosis factor alpha; IL-6, interleukin 6; TBARS,
thiobarbituric acid reactive substances.
In a randomized, controlled, cross-over trial involving 30 patients with T2DM
and 6 patients with impaired glucose tolerance, patients took either 900 mL/d of
HRW or 900 mL of placebo water for 8 weeks, with a 12-week period of washout
[74]. Intake of HRW led to significant declines in modified LDL-C
(i.e., modifications that increase the net negative charge of LDL), small dense
LDL, and urinary 8-isoprostanes by 15.5% (p 0.01), 5.7%
(p 0.05), and 6.6% (p 0.05), respectively [74].
Additionally, there was a trend towards lower serum concentrations of oxidized
LDL and free fatty acids, as well as increased plasma concentrations of
adiponectin and extracellular SOD [74]. These results suggest that HRW could be a
useful adjunct in preventing T2DM and insulin resistance, potentially by
activating ATP-binding cassette transporter A1-dependent efflux and enhancing the
anti-atherosclerotic functions of HDL, and have beneficial lipid-lowering effects
[74]. These findings suggest that HRW could be a useful adjunct in preventing
T2DM and insulin resistance, potentially by activating ATP-binding cassette
transporter A1-dependent cholesterol efflux and enhancing the
anti-atherosclerotic functions of HDL [74]. Since MS has become a worldwide
problem, H therapy may be a new approach for mitigating CMDs [73, 74, 75, 76, 77]. A
recent review has also reemphasized that Indo-Mediterranean diets can produce
greater H, and may be a better option for preventing hypertension [78].
H therapy may have a beneficial impact on mitochondrial function, as shown
in a rat study conducted by Gvozdjáková et al. [79]. The study
showed that administering H to rats led to enhanced state 3 respiration in
cardiac mitochondria, linked to both Complex I (CI) and Complex II (CII)
substrates [79]. It was proposed that H may facilitate the conversion of
quinone intermediates in the Q-cycle to the fully reduced ubiquinol [80]. This
conversion could boost the antioxidant capacity of the quinone pool, thereby
reducing the generation of mitochondrial ROS [80].
11. Conclusions
The past two decades have seen increased interest in the potential health
benefits of H, particularly in cardiovascular and metabolic diseases. The
primary mechanism behind H’s therapeutic effects appears to be its
selective and efficient scavenging and neutralization of ROS such as
•OH and •ONOO. Beyond its antioxidant role, H
also exhibits anti-inflammatory and anti-apoptotic properties. Our review
indicates that H administration shows promise in mitigating CVDs,
atherosclerosis, stroke, and hyperlipidemia, with potential applicability in
coronary artery disease and diabetes. Notably, H can be endogenously produced in
the human gut by specific bacteria, a process that can be optimized through
dietary choices. For example, a Mediterranean-style diet, rich in fiber and
bioactive compounds, may enhance gut-based H production. Our review indicates
that H administration shows promise in mitigating CVDs, atherosclerosis,
stroke, and hyperlipidemia, with potential applicability in coronary artery
disease and diabetes. Notably H may be produced in gut by bacteria in the
human body. This process can be optimized through dietary choices, particularly
the MD which is rich in fiber and bioactive compounds. Given the growing body of
evidence supporting H’s positive impact on metabolic and cardiovascular
conditions, targeted strategies to increase intestinal H production may
serve as future preventive measures or adjunctive treatments of these diseases.
Author Contributions
The first draft of the manuscript was made by RBS and VM which was critically
reviewed by ZS, JF, GF, VM, AT, OP, AG, KF, JV, BKur, BKal, JS. RBS, VM, JS, BKur
and BKal made substantial contributions to conception and design of the
manuscript, ZS, GF, JF, AT, OP, AG, KF, PZ, and JV participated in the
visualization, editing, and funding acquisition of the manuscript. All coauthors
made critical comments which were incorporated in the article and agreed to be
accountable for all aspects of the work. All authors read and approved the final
manuscript.
Ethics Approval and Consent to Participate
Not applicable.
Acknowledgment
Not applicable.
Funding
This work was supported by grants from Slovak Research and Development Agency
(APVV-15-0376, APVV-19-0317), grant from the Slovak Academy of Sciences (VEGA
2/0092/22, 2/0063/18 and 2/0148/22), and grant from European Union Structural
funds (ITMS 26230120009), grant (2018/7838:1-26C0), grant from Ministry of Health
of The Slovak Republic (2019/4-CEMSAV-1) and grant (Biovid: ITMS2014+:
313011AVG3).
Conflict of Interest
The authors declare no conflict of interest. Jan Slezak is serving as one of the Editorial Board members of this journal. We declare that Jan Slezak had no involvement in the peer review of this article and has no access to information regarding its peer review. Full responsibility for the editorial process for this article was delegated to Vincenzo Lionetti.