Effects Of Hydrogen Inhalation Research Articles

Effect of hydrogen inhalation on cardiovascular and interstitial components of pulmonary hypertension in rats

Hydrogen is known to have selective antioxidant properties. It binds highly reactive hydroxyl radicals. The pathogenesis of the monocrotaline animal model of pulmonary hypertension is associated with oxidative stress and leads to all the symptoms of pulmonary hypertension (PH) and interstitial lung disease (ILD) associated with hypertension.The aim of this work was to study the effect of 4% hydrogen inhalations on the symptoms of PH and ILD in rats.Methods. To model monocrotaline-induced pulmonary hypertension (MCT-PH), two groups of animals received a single subcutaneous injection of monocrotaline (MCT) on day 1. The control group was injected subcutaneously with MCT solvent only. The animals receiving MCT were further divided into 2 subgroups. Subgroup 1 rats breathed room air and subgroup 2 rats breathed a mixture of room air and 4% hydrogen. The regular inhalations continued until day 21. On day 21, hemodynamic parameters were measured under urethane anesthesia, the heart and its components and the lungs were weighed, and lung tissue was preserved for morphological study.Results. The inhalation had no effect on the main cardiovascular symptoms of PH, but a positive effect on the state of the connective tissue of the lungs affected by PH was shown. The mast cell response was reduced both quantitatively and functionally. There was a decrease in tryptase expression by mast cells, with predominance of the forms without signs of degranulation. TGF-β secretion was also significantly reduced and visualized by immunopositive cells in alveolar cellular structures and vessel walls.Conclusion. Inhalation of 4% hydrogen reduces inflammation and fibrosis of lung tissue during the development of MCT-PH.

Effect of hydrogen inhalation on mitochondrial biosynthesis in lung tissues during acute lung injury in mice with sepsis

Objective To evaluate the effect of hydrogen(H2)inhalation on mitochondrial biosynthesis in lung tissues during acute lung injury in mice with sepsis. Methods One hundred and four male ICR mice, aged 6 weeks, weighing 20-25 g, were divided into 4 groups(n=26 each)using a random number table: sham operation group(group S), sham operation + H2 group(group S+ H2), sepsis group(group Sep)and sepsis + H2 group(group Sep+ H2). Sepsis was produced by cecal ligation and puncture.In S+ H2 and Sep+ H2 groups, the mice inhaled 2% H2 for 1 h starting from 1 and 6 h after operation.Twenty mice in each group were selected, and the survival rates on postoperative days 1, 2, 3, 5 and 7 were recorded.On the postoperative day 1, 6 mice in each group were selected, and blood samples were collected from the common carotid artery for measurement of arterial oxygen partial pressure, and the oxygenation index was calculated.The pulmonary specimens were obtained for examination of the pathological changes which were scored and for determination of the expression of peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)in lung tissues by Western blot.The pulmonary mitochondria were isolated for determination of mitochndrial membrane potential(MMP)and ATP contents using spectrophotometry and a bioluminescence technique, respectively. Results Compared with group S, the survival rate, oxygenation index and MMP and ATP content in lung tissues were significantly decreased, and the pathological scores and PGC-1α expression in lung tissues were significantly increased in Sep and Sep+ H2 groups(P<0.05). Compared with group Sep, the survival rate, oxygenation index, and MMP, ATP content and PGC-1α expression in lung tissues were significantly increased, and the pathological scores were significantly decreased in group Sep+ H2(P<0.05). Conclusion H2 inhalation can ameliorate acute lung injury in mice with sepsis, and the mechanism is associated with the enhanced function of PGC-1α and promoted mitochondrial biosynthesis in lung tissues. Key words: Hydrogen; Sepsis; Respiratory distress syndrome, adult; Mitochondria

Effects of hydrogen inhalation on serum pro-inflammatory factors and intestinal injury in mice with severe sepsis

To investigate the effects and mechanisms of hydrogen inhalation on serum levels of pro-inflammatory factors and intestinal injury in severe septic mice. 176 male ICR mice were randomly divided into four groups: sham operation group, hydrogen control group (sham+hydrogen inhalation), model group (severe sepsis model) and hydrogen treatment group (severe sepsis model+hydrogen inhalation), with 44 mice in each group. Severe sepsis model was reproduced by cecal ligation and puncture (CLP). 2% hydrogen inhalation was given for 1 hour at 1 hour and 6 hours after sham or CLP operation. Twenty animals in each group were selected and observed for 7-day survival rate. Six animals in each group were selected and sacrificed at 6, 12, 24 and 48 hours after sham or CLP, the concentrations of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-10) and high mobility group box 1 (HMGB1) in serum were determined, the intestinal histopathological changes and scores were evaluated by light microscopy, and the activities of myeloperoxidase (MPO) and caspase-3 were determined. The 7-day survival rate of severe sepsis mice was 0; the 7-day survival rate was increased to 60% in hydrogen treatment group, with statistical significance in variables compared with model group (P<0.05). Compared with sham operation group, the serum concentrations of TNF-α, IL-6, IL-10 and HMGB1 were obviously increased, the intestine were heavily injured along with higher histopathological scores, and the intestinal MPO and caspase-3 activities were significantly enhanced at different time points after CLP in model group (all P<0.05). Compared with model group, the serum concentrations of TNF-α, IL-6 and HMGB1 were significantly decreased [ TNF-α (ng/L): 6 hours: 110.34±9.28 vs. 440.55±25.78, 12 hours: 82.29±8.43 vs. 448.36±32.54, 24 hours: 79.68±9.04 vs. 346.42±22.24, 48 hours: 80.79±10.06 vs. 368.94±31.58; IL-6 (ng/L): 12 hours: 58.68±8.55 vs. 158.28±16.73, 24 hours: 46.98±7.58 vs. 146.74±18.02, 48 hours: 38.67±8.22 vs. 136.45±15.45; HMGB1 (μg/L): 6 hours: 15.75±4.32 vs. 55.56±10.04, 12 hours: 32.02±9.33 vs. 89.65±15.65, 24 hours: 35.87±8.54 vs. 86.44±20.33, 48 hours: 23.85±9.83 vs. 98.33±18.88, all P<0.05], the serum concentrations of IL-10 (ng/L) at 24 hours and 48 hours after CLP were obviously increased (24 hours: 135.44±16.43 vs. 79.22±12.03, 48 hours: 110.92±12.54 vs. 74.47±11.18, both P<0.05), the intestinal injury were ameliorated with decreased histopathological scores (12 hours: 1.70±0.06 vs. 3.23±0.44, 24 hours: 2.12±0.31 vs. 4.51±0.58, 48 hours: 2.03±0.42 vs. 4.27±0.58, all P<0.05), and the intestinal MPO and caspase-3 activities were significantly decreased [MPO (U/g): 6 hours: 13.75±4.21 vs. 25.56±5.34, 12 hours: 14.72±4.22 vs. 30.53±6.87, 24 hours: 11.62±3.14 vs. 33.58±7.24, 48 hours: 11.33±4.03 vs. 38.57±8.12; caspase-3 (fluorescence intensity): 6 hours: 0.37±0.07 vs. 0.69±0.23, 12 hours: 0.42±0.07 vs. 0.86±0.13, 24 hours: 0.53±0.11 vs. 1.36±0.23, 48 hours: 0.50±0.08 vs. 1.48±0.15, all P<0.05] in hydrogen treatment group. Hydrogen inhalation can down-regulate the systemic inflammatory response to ameliorate the intestinal injury, and it may improve the septic process and increase the survival rate of mice with severe sepsis.

Relieving eff ect of hydrogen inhalation to donor lung on posttransplantational ischemia-reperfusion injury

Relieving eff ect of hydrogen inhalation to donor lung on posttransplantational ischemia-reperfusion injury

Effect of hydrogen inhalation on brain injury in septic mice

Objective To evaluate the effect of hydrogen (H2) inhalation on brain injury in septic mice.Methods One hundred and eighty male ICR mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 4 groups (n =45 each) using a random number table:sham operation group (SH group),H2 group,sepsis group (S group) and sepsis + H2 group (S + H2 group).Sepsis was produced by cecal ligation and puncture (CLP) in chloral hydrate-anesthetized mice.H2 and S + H2 groups inhaled 2％ H2 for 1 h starting from 1 and 6 h after CLP.At 7,12 and 24 h after CLP,6 mice in each group were sacrificed and brains were removed for determination of Evans Blue (EB) and water contents in brain tissues.Another 6 mice in each group were chosen at 7,12 and 24 h after CLP,blood samples were obtained,the mice were then sacrificed and brains were removed to measure the concentrations of tumor necrosis factor-α (TNF-α),high mobility group protein B1 (HMGB1) and interleukin-10 (IL-10) in serum and brain tissues by ELISA.Three mice in each group were sacrificed at 24 h after LPS and brains were removed for microscopic examination of pathological changes in hippocampal CA1 region.Six mice in each group were sacrificed at 24 h after LPS and brains were removed for determination of the expression of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in brain tissues by Western blot.Results Compared with SH group,the contents of EB and water in brain tissues were significantly increased,and the levels of TNF-α,HMGB1 and IL-10 in serum and brain tissues were increased at 7,12 and 24 h after CLP,and the expression of Nrf2 and HO-1 in brain tissues was up-regulated at 24 h after CLP in S and CLP + H2 groups,and no significant difference was found in the index above in H2 group.Compared with S group,the contents of EB and water in brain tissues were significantly decreased,the levels of TNF-α and HMGB1 in serum and brain tissues were decreased and the level of IL-10 in serum and brain tissues was increased at 7,12 and 24 h after CLP,and the expression of Nrf2 and HO-1 in brain tissues was up-regulated at 24 h after CLP and the pathological changes were attenuated in group CLP + H2.Conclusion H2 inhalation can attenuate brain injury in septic mice and the mechanism is related to activation of Nrf2/ARE signaling pathway and inhibition of inflammatory responses. Key words: Hydrogen ; Sepsis ; Brain injuries

Effect of hydrogen inhalation on lung ischemia-reperfusion injury in mice

Objective To investigate the effect of hydrogen inhalation on lung ischemia-reperfusion (I/R) injury in mice.Methods Fifty male C3H/HeN mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 5 groups (n =10 each) using a random number table:sham operation group (group SH),I/R group,1％ H2 group,2％ H2 group and 4％ H2 group.Lung I/R was produced by occlusion of the left hilum of lungs for 60 min followed by 240 min reperfusion.1％,2％ and 4％ H2 were inhaled during reperfusion in 1％ H2,2％ H2 and 4％ H2 groups,respectively.At the end of reperfusion,blood samples were obtained for blood gas analysis.PaO2/FiO2 was calculated.The animals were then sacrificed and lungs were immediately removed for determination of the levels of superoxide dismutase (SOD),malonaldehyde (MDA),tumor necrosis factor-α (TNF-α),interleukin-8 (IL-8),Toll-like receptor 4 (TLR4) and myeloperoxidase (MPO) in the lung tissues and for microscopic examination of the pathological changes of lungs which were scored.Results Compared with SH group,PaO2/FiO2 was significantly decreased,the pathological scores,levels of MDA,TNF-α,IL-8,TLR4 and MPO were increased,and SOD activity was decreased in I/R,1％ H2,2％ H2 and 4％ H2 groups.Compared with group I/R,PaO2/FiO2 was significantly increased,the pathological scores and levels of MDA,TNF-α,IL-8,TLR4 and MPO in lung tissues were decreased,and SOD activity was increased in 1％ H2,2％ H2 and 4％ H2 groups.There were no significant differences in the indices mentioned above among 1％ H2,2％ H2 and 4％ H2 groups.Conclusion Hydrogen inhalation can alleviate lung I/R injury in mice,and the mechanism is related to inhibition of lipid peroxidation responses and inflammatory responses. Key words: Hydrogen ; Reperfusion injury ; Lung

(115) - The Effects of Hydrogen Inhalation on Non-Beating Heart Donor Lung During Ex Vivo Lung Perfusion

Lung transplantation (LTx) is an effective treatment for end-stage lung disease. However, the shortage of donor lung causes the death of recipients in waiting list. In order to resolve the shortage of donor lung, non-beating heart donor (NBHD) lungs are used and ex vivo lung perfusion (EVLP) is performed in clinical setting. In this study, the effects of hydrogen gas known as antioxidant and anti-inflammatory gas on NBHD lungs were investigated during EVLP. Ten 40kg female pigs were randomly divided into control (group I, n=5) and study group (group II, n=5). The atrial fibrillation was induced by 9V electric shock and one hour of warm ischemic injury was induced by leaving them without any treatments after cardiac arrest. After flush using Perfadex solution, the harvested lungs were applied to EVLP system for 4 hours. Steen solution was used as perfusate. During EVLP, the lungs were ventilated with room air (0.2 of FiO2) in control group and 2% hydrogen gas in study group. Oxygen capacity ((PaO2- pulmonary artery of the left atrium PaO2) / FiO2), pulmonary vascular resistance (Pulmonary artery pressure-left atrium pressure) X 80/ pulmonary artery flow) and peak airway pressure were measured every hour for 4 hours of EVLP. After the EVLP, lung specimens were excised for measurement of the extent of p38, JNK, and ERK 1/2 phosphorylation and the expression of IL-1ß, IL-6, IL-8, and TNF-α. Pathologic evaluation was performed using lung injury severity score after H & E stain. Wet/dry ratio was examined. Oxygen capacity in study group was higher than in control group, which did not show a significant difference (p = 0.0862). However, pulmonary vascular resistance (p = 0.0111) and peak airway pressure (p = 0.0189) were significantly lower in study group. There was a tendency that the extents of phosphorylation of p38, JNK, and ERK 1/2 were lower in study group, but it was not statistically significant. The expressions of IL-1ß (p = 0.0317), IL-6 (p = 0.0159), IL-8 (p = 0.0195), and TNF-α (p = 0.0159) were all significantly lower in study group. Lung injury score (p = 0.0358) and wet/dry ratio (p = 0.040) were also lower in study group. The inhalation of hydrogen gas during EVLP improved the function of NBHD lung and it might increase the utilization of NHBD lung with warm ischemic injury.

The protective effects of hydrogen inhalation on renal injury in severe septic mice

Objective To investigate the protective effects of hydrogen gas (H2) inhalation on acute renal injury in severe septic mice,and the associated mechanism.Methods Sepsis model was made by cecal ligation and puncture (CLP) operation in mice.A total of 24 mice were randomly (random number)divided into four groups:sham operation group (Sham),sham operation with H2 inhalation group (Sham +H2),sepsis group (Sepsis),and sepsis with H2 inhalation group (Sepsis + H2).The procedure for inhalation of 2％ H2 for 1 h was initiated at 1 h after CLP and 6 h after sham operation,respectively.The pathological changes and apoptosis of kidney,the blood creatinine (Cr) and blood urea nitrogen (BUN)concentration,the activities of superoxide dimutase (SOD) and catalase (CAT) in serum and kidney,as well as the levels of 8-iso-prostaglandin F2α (8-iso-PGF2α) and high mobility group box 1 (HMGB1) in serum and kidney were observed at 24 h after operation.Results H2 inhalation could significantly alleviate the renal injury of septic mice,reduce the renal cell apoptosis,increase the activities of SOD and CAT,as well as decrease the levels of 8-iso-PGF2α and HMGB1 in serum and kidney (P ＜ 0.05).Conclusions H2 inhalation can alleviate acute renal injury in septic mice,resulting from the increased activities of antioxidant enzymes and the reduced levels of oxidative products and inflammatory cytokines. Key words: Hydrogen ; Sepsis ; Acute renal injury ; Antioxidant enzyme ; Inflammatory cytokine