Inhalation of hydrogen gas reduces infarct size in the rat model of myocardial ischemia–reperfusion injury

doi:10.1016/j.bbrc.2008.05.165

Biochemical and Biophysical Research Communications

Volume 373, Issue 1, 15 August 2008, Pages 30-35

https://doi.org/10.1016/j.bbrc.2008.05.165 Get rights and content

Abstract

Inhalation of hydrogen (H₂) gas has been demonstrated to limit the infarct volume of brain and liver by reducing ischemia–reperfusion injury in rodents. When translated into clinical practice, this therapy must be most frequently applied in the treatment of patients with acute myocardial infarction, since angioplastic recanalization of infarct-related occluded coronary artery is routinely performed. Therefore, we investigate whether H₂ gas confers cardioprotection against ischemia–reperfusion injury in rats. In isolated perfused hearts, H₂ gas enhances the recovery of left ventricular function following anoxia-reoxygenation. Inhaled H₂ gas is rapidly transported and can reach ‘at risk’ ischemic myocardium before coronary blood flow of the occluded infarct-related artery is reestablished. Inhalation of H₂ gas at incombustible levels during ischemia and reperfusion reduces infarct size without altering hemodynamic parameters, thereby preventing deleterious left ventricular remodeling. Thus, inhalation of H₂ gas is promising strategy to alleviate ischemia–reperfusion injury coincident with recanalization of coronary artery.

Section snippets

Materials and methods

Animals. All experimental procedures and protocols were approved by the Animal Care and Use Committees of the Keio University and conformed to the NIH Guide for the Care and Use of Laboratory Animals. Eight-week-old male Wistar rats were artificially ventilated under anesthesia with ketamine (60 mg/kg) and xylazine (15 mg/kg) given intraperitoneally. Temperature was maintained at 37.5 ± 0.5 °C using a thermostatically controlled heating blanket connected to a thermometer probe placed in the rectum. H

H₂ gas improves the recovery of left ventricular function during reoxygenation after anoxia in isolated perfused hearts

We first studied the effect of H₂ gas on the functional recovery after anoxia-reoxygenation in Langendorff-perfused rat hearts. Hearts were subjected to 40 min of anoxic perfusion with buffer equilibrated with either 100% N₂ (Control group) or 100% H₂ (H₂ group) followed by 40 min of aerobic reperfusion with buffer equilibrated with 95% O₂ and 5% CO₂ (Fig. 1A). H₂ gas significantly improved the recovery of LV developed pressure (LVDP), positive dP/dt, and negative dP/dt 40 min after reoxygenation (

Discussion

This is the first study to demonstrate that inhalation of H₂ gas, at an incombustible level, limit the extent of myocardial infarction resulting from myocardial ischemia–reperfusion injury, and thereby preserve LV function in vivo. The cardioprotective effect of H₂ gas was also confirmed ex vivo Langendorff-perfused hearts subjected to anoxia-reoxygenation injury. The anti-oxidant properties of H₂ were confirmed by the demonstration that (1) H₂ improves the recovery of LV function during

Acknowledgments

We thank M. Okada (NIHON KODEN), S. Kotouda (LMS laboratory and Medical Supplies), C. Ogawa, K. Nishimaki, M. Kamimura, M, Abe, Y. Miyake, H. Kawaguchi, H. Shiozawa, and M. Ono for their technical assistance. M. Sano is a core member of the Global Center-of-Excellence (GCOE) for Human Metabolomics Systems Biology from MEXT. This work was supported by a PRESTO (Metabolism and Cellular Function) grant from the Japanese Science and Technology Agency awarded to M. Sano.

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Cited by (415)

Protective Effects of Hydrogen Gas Inhalation for Hindlimb Ischaemia–Reperfusion Injury in a Mouse Model
2024, European Journal of Vascular and Endovascular Surgery
Ischaemia–reperfusion (I/R) injury is a severe post-operative complication that triggers an inflammatory response and causes severe damage. Hydrogen gas has anti-oxidant and anti-apoptotic properties and has been shown to be safe in humans. The study aimed to investigate whether hydrogen gas protects against skeletal muscle I/R injury.
Experimental basic research using mice. A total of 160 eight to 10 week old albino laboratory bred strain of house mice (25.8 ± 0.68 g) were used in this study. The mice were cable tied to the hindlimb under anaesthesia and then placed in an anaesthesia box filled with air and 2% isoflurane (control group); 80 mice were additionally subjected to 1.3% hydrogen gas in this mix (hydrogen group). After two hours, the cable ties were removed to initiate reperfusion, and hydrogen inhalation lasted for six hours in the hydrogen group. After six hours, the mice were taken out of the box and kept in cages under standard conditions until time for observation at 16 different time points after reperfusion: zero, two, four, six, eight, and 10 hours and one, two, three, four, five, six, seven, 14, 21, and 28 days. Five mice were sacrificed using excess anaesthesia at each time point, and the bilateral hindlimb tissues were harvested. The inflammatory effects of the I/R injury were assessed by evaluating serum interleukin-6 concentrations using enzyme linked immunosorbent assay, as well as histological and immunohistochemical analyses. Untreated mice with I/R injury were used as controls.
Hydrogen gas showed protective effects associated with a reduction in inflammatory cell infiltration (neutrophils, macrophages, and lymphocytes), a reduced area of damaged muscle, maintenance of normal muscle cells, and replacement of damaged muscle cells with neoplastic myocytes.
Inhalation of hydrogen gas had a protective effect against hindlimb I/R injury in mice, in part by reducing inflammatory cell infiltration and in part by preserving normal muscle cells.
Protective effects of hydrogen rich saline solution in rats with experimental myocardial ischemia reperfusion injury
2023, Heliyon
The aim of our study is to show whether the administration of hydrogen-rich saline solution (HRSS) intraperitoneally before left main coronary artery (LAD) ischemia protects the myocardium against ischemia-reperfusion (IR) injury.
After ethics committee approval, 24 Wistar Albino rats were divided into 4 groups, 6 rats in each group. For experimental IR, myocardial ischemia was performed by LAD ligation. Left thoracotomy was performed without ischemia in the Control group (Group C). Left thoracotomy was performed without myocardial ischemia to the rats in the HRSS group, and HRSS was given intraperitoneally (ip) at a rate of 10 ml/kg throughout the procedure. In the MIR-HRSS group, a single dose of 10 ml/kg HRSS was administered 5 min before reperfusion. Histopathological and biochemical parameters were compared in myocardial tissue samples taken at the end of the reperfusion period.
When the groups were compared among themselves in terms of TOS and TAS levels, there was a significant difference between the groups (p = 0.006, p = 0.002). The severity of cardiomyocyte degeneration was significantly greater in MIR group than that in the control and HRSS groups (p = 0.002 and p = 0.001, respectively), as well as severity score of cardiomyocyte degeneration was higher in MIR-HRSS group compared with HRSS group (p = 0.035).
Our study shows that HRSS is protective in IR injury, with the application of HRSS 5 min before reperfusion, interstitial edema severity, subendocardial haemorrhage are reduced, and oxidant status parameters are increased, while antioxidant status parameters are decreased. We believe that when it is supported by other studies, the protective effects of HRSS on IR damage will be shown in detail and its indications will be expanded.
Efficacy of inhaled hydrogen on neurological outcome following brain ischaemia during post-cardiac arrest care (HYBRID II): a multi-centre, randomised, double-blind, placebo-controlled trial
2023, eClinicalMedicine
Inhaled molecular hydrogen gas (H₂) has been shown to improve outcomes in animal models of cardiac arrest (CA). H₂ inhalation is safe and feasible in patients after CA. We investigated whether inhaled H₂ would improve outcomes after out-of-hospital CA (OHCA).
HYBRID II is a prospective, multicentre, randomised, double-blind, placebo-controlled trial performed at 15 hospitals in Japan, between February 1, 2017, and September 30, 2021. Patients aged 20–80 years with coma following cardiogenic OHCA were randomly assigned (1:1) using blinded gas cylinders to receive supplementary oxygen with 2% H₂ or oxygen (control) for 18 h. The primary outcome was the proportion of patients with a 90-day Cerebral Performance Category (CPC) of 1 or 2 assessed in a full-analysis set. Secondary outcomes included the 90-day score on a modified Rankin scale (mRS) and survival. HYBRID II was registered with the University Hospital Medical Information Network (registration number: UMIN000019820) and re-registered with the Japan Registry for Clinical Trials (registration number: jRCTs031180352).
The trial was terminated prematurely because of the restrictions imposed on enrolment during the COVID-19 pandemic. Between February 1, 2017, and September 30, 2021, 429 patients were screened for eligibility, of whom 73 were randomly assigned to H₂ (n = 39) or control (n = 34) groups. The primary outcome, i.e., a CPC of 1 or 2 at 90 days, was achieved in 22 (56%) and 13 (39%) patients in the H₂ and control groups (relative risk compared with the control group, 0.72; 95% CI, 0.46–1.13; P = 0.15), respectively. Regarding the secondary outcomes, median mRS was 1 (IQR: 0–5) and 5 (1–6) in the H₂ and control groups, respectively (P = 0.01). An mRS score of 0 was achieved in 18 (46%) and 7 (21%) patients in the H₂ and control groups, respectively (P = 0.03). The 90-day survival rate was 85% (33/39) and 61% (20/33) in the H₂ and control groups, respectively (P = 0.02).
The increase in participants with good neurological outcomes following post-OHCA H₂ inhalation in a selected population of patients was not statistically significant. However, the secondary outcomes suggest that H₂ inhalation may increase 90-day survival without neurological deficits.
Taiyo Nippon Sanso Corporation.
For the Japanese translation of the abstract see Supplementary Materials section.
Taurine, Coenzyme Q<inf>10</inf>, and Hydrogen Water Prevents Germanium Dioxide-Induced Mitochondrial Dysfunction and Associated Sensorineural Hearing Loss in mouse
2023, Hearing Research
Mitochondrial dysfunction has been implicated in numerous common diseases as well as aging and plays an important role in the pathogenesis of sensorineural hearing loss (SNHL). In the current study, we showed that supplementation with germanium dioxide (GeO₂) in CBA/J mice resulted in SNHL due to the degeneration of the stria vascularis and spiral ganglion, which were associated with down-regulation of mitochondrial respiratory chain associated genes and up-regulation in apoptosis associated genes in the cochlea. Supplementation with taurine, coenzyme Q10, or hydrogen-rich water, attenuated the cochlear degeneration and associated SNHL induced by GeO₂. These results suggest that daily supplements or consumption of antioxidants, such as taurine, coenzyme Q10, and hydrogen-rich water, may be a promising intervention to slow SNHL associated with mitochondrial dysfunction.
E3 ubiquitin ligase COP1 confers neuroprotection in cerebral ischemia/reperfusion injury via regulation of transcription factor C/EBPβ in microglia
2022, International Journal of Biological Macromolecules
This study intends to investigate the potential involvement of E3 ubiquitin ligase COP1 in cerebral ischemia-reperfusion (I/R) injury.
A mouse model of middle cerebral artery occlusion/reperfusion (MCAO/R) was established, and the ischemic penumbra of mouse brain cortex was collected and subjected to RNA-sequencing (RNA-seq). Primary glial cells, neurons and astrocytes were isolated, and microglia were exposed to oxygen and glucose deprivation/reperfusion (OGD/R).
COP1 was poorly expressed in MCAO mice and OGD/R microglia. Restoration of COP1 suppressed the activation of microglia and relieves neuroinflammation in cerebral I/R injury, leading to alleviated brain damage (infraction volume [%]: [31.58 ± 2.96] & [12.06 ± 1.29], neurological scores: [3.6 ± 0.6] & [1.2 ± 0.5]). COP1 promoted the ubiquitin-mediated degradation of C/EBPβ in microglia. It was further revealed that COP1 attenuated microglia activation and phagocytosis (Iba + cells [N/mm²]: 182.68 ± 20.89 & 84.57 ± 12.08; soma area [μm²]: 78.24 ± 8.75 & 59.78 ± 7.61) through negative regulation of C/EBPβ protein expression. Restoration of C/EBPβnegated the neuroprotective effects of COP1 in vivo.
This study illuminated a mechanism by which COP1 conferred a neuroprotective role in cerebral I/R injury via enhancing the ubiquitin-mediated degradation of transcriptional factor C/EBPβ in microglia.
Protective effects of hydrogen gas against spinal cord ischemia–reperfusion injury
2022, Journal of Thoracic and Cardiovascular Surgery
This experimental study aimed to assess the efficacy of hydrogen gas inhalation against spinal cord ischemia–reperfusion injury and reveal its mechanism by measuring glutamate concentration in the ventral horn using an in vivo microdialysis method.
Male Sprague-Dawley rats were divided into the following 6 groups: sham, only spinal ischemia, 3% hydrogen gas (spinal ischemia + 3% hydrogen gas), 2% hydrogen gas (spinal ischemia + 2% hydrogen gas), 1% hydrogen gas (spinal ischemia + 1% hydrogen gas), and hydrogen gas dihydrokainate (spinal ischemia + dihydrokainate [selective inhibitor of glutamate transporter-1] + 3% hydrogen gas). Hydrogen gas inhalation was initiated 10 minutes before the ischemia. For the hydrogen gas dihydrokainate group, glutamate transporter-1 inhibitor was administered 20 minutes before the ischemia. Immunofluorescence was performed to assess the expression of glutamate transporter-1 in the ventral horn.
The increase in extracellular glutamate induced by spinal ischemia was significantly suppressed by 3% hydrogen gas inhalation (P < .05). This effect was produced in increasing order: 1%, 2%, and 3%. Conversely, the preadministration of glutamate transporter-1 inhibitor diminished the suppression of spinal ischemia-induced glutamate increase observed during the inhalation of 3% hydrogen gas. Immunofluorescence indicated the expression of glutamate transporter-1 in the spinal ischemia group was significantly decreased compared with the sham group, which was attenuated by 3% hydrogen gas inhalation (P < .05).
Our study demonstrated hydrogen gas inhalation exhibits a protective and concentration-dependent effect against spinal ischemic injury, and glutamate transporter-1 has an important role in the protective effects against spinal cord injury.

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Biochemical and Biophysical Research Communications

Abstract

Section snippets

Materials and methods

H₂ gas improves the recovery of left ventricular function during reoxygenation after anoxia in isolated perfused hearts

Discussion

Acknowledgments

References (24)

Primary angioplasty versus intravenous thrombolytic therapy for acute myocardial infarction: a quantitative review of 23 randomised trials

Lancet

Measurement of superoxide-derived free radicals in the reperfused heart. Evidence for a free radical mechanism of reperfusion injury

J. Biol. Chem.

Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress

Biochem. Biophys. Res. Commun.

Short-term caloric restriction improves ischemic tolerance independent of opening of ATP-sensitive K⁺ channels in both young and aged hearts

J. Mol. Cell. Cardiol.

Aging impairs functional, metabolic and ionic recovery from ischemia–reperfusion and hypoxia-reoxygenation

J. Mol. Cell. Cardiol.

Hydrogenase

Biochim. Biophys. Acta

Determinants of 6-month mortality in survivors of myocardial infarction after thrombolysis. Results of the GISSI-2 data base. The Ad hoc Working Group of the Gruppo Italiano per lo Studio della Sopravvivenza nell’Infarto Miocardico (GISSI)-2 Data Base

Circulation

Myocardial reperfusion: a double-edged sword?

J. Clin. Invest.

Myocardial reperfusion injury

N. Engl. J. Med.

Demonstration of free radical generation in “stunned” myocardium of intact dogs with the use of the spin trap alpha-phenyl N-tert-butyl nitrone

J. Clin. Invest.

Preconditioning in cardiomyocytes protects by attenuating oxidant stress at reperfusion

Circ. Res.

Recombinant human superoxide dismutase (h-SOD) fails to improve recovery of ventricular function in patients undergoing coronary angioplasty for acute myocardial infarction

Circulation

Cited by (415)

Protective Effects of Hydrogen Gas Inhalation for Hindlimb Ischaemia–Reperfusion Injury in a Mouse Model

Protective effects of hydrogen rich saline solution in rats with experimental myocardial ischemia reperfusion injury

Efficacy of inhaled hydrogen on neurological outcome following brain ischaemia during post-cardiac arrest care (HYBRID II): a multi-centre, randomised, double-blind, placebo-controlled trial

Taurine, Coenzyme Q<inf>10</inf>, and Hydrogen Water Prevents Germanium Dioxide-Induced Mitochondrial Dysfunction and Associated Sensorineural Hearing Loss in mouse

E3 ubiquitin ligase COP1 confers neuroprotection in cerebral ischemia/reperfusion injury via regulation of transcription factor C/EBPβ in microglia

Protective effects of hydrogen gas against spinal cord ischemia–reperfusion injury

Inhalation of hydrogen gas reduces infarct size in the rat model of myocardial ischemia–reperfusion injury

Abstract

Section snippets

Materials and methods

H2 gas improves the recovery of left ventricular function during reoxygenation after anoxia in isolated perfused hearts

Discussion

Acknowledgments

Lancet

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Mol. Cell. Cardiol.

J. Mol. Cell. Cardiol.

Biochim. Biophys. Acta

Determinants of 6-month mortality in survivors of myocardial infarction after thrombolysis. Results of the GISSI-2 data base. The Ad hoc Working Group of the Gruppo Italiano per lo Studio della Sopravvivenza nell’Infarto Miocardico (GISSI)-2 Data Base

Circulation

Myocardial reperfusion: a double-edged sword?

J. Clin. Invest.

Myocardial reperfusion injury

N. Engl. J. Med.

Demonstration of free radical generation in “stunned” myocardium of intact dogs with the use of the spin trap alpha-phenyl N-tert-butyl nitrone

J. Clin. Invest.

Preconditioning in cardiomyocytes protects by attenuating oxidant stress at reperfusion

Circ. Res.

Recombinant human superoxide dismutase (h-SOD) fails to improve recovery of ventricular function in patients undergoing coronary angioplasty for acute myocardial infarction

Circulation

H₂ gas improves the recovery of left ventricular function during reoxygenation after anoxia in isolated perfused hearts