Effects Of Hydrogen-rich Saline Research Articles

Effects of hydrogen-rich saline on incisional pain-remifentanil-induced hyperalgesia in rats

Objective To evaluate the effects of hydrogen-rich saline on incisional pain-remifentanil-induced hyperalgesia in rats.Methods Thirty-two male Sprague-Dawley rats,aged 2-3 months,weighing 240-260 g,in which the catheter was successfully inserted into the caudal vein,were randomly divided into 4 groups (n =8 each) using a random number table:control gourp (group C),remifentanil + incisional pain group (group R + I) and different doses of hydrogen-rich saline groups (H1 and H2 groups).A l-cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the left hindpaw in chloral hydrate-anesthetized rats.Remifentanil 1 μg· kg-1 · min-1 was infused intravenously for 60 min sarting from beginning of establishment of incisional pain model in R + I,H1 and H2 groups.The equal volume of normal saline was infused intravenously for 60 rin instead of remifentanil group C.Hydrogen-rich saline 5 and 10 ml/kg were injected intraperitoneally at 10 min before establishment of incisional pain model in H1 and H2 groups,respectively.Paw withdrawal threshold (PWT) to yon Frey hair stimulation and paw withdrawal latency (PWL) to thermal stimulation were measured at 24 h before remifentanil infusion and 2,6,24 and 48 h after remifentanil infusion (T0-T4).The rats were sacrificed after measuremnt of pain threshold,and L4-6 segments of the spinal cord were removed for determination of the expression of R1 and 2B subunits-containing N-methyl-D-aspartate receptors (NR1 and NR2B) in total and membrane proteins by Western blot.The ratio between the expression of NR1 in membrane protein and in total protein (mNR1/tNR1) and NR2B in membrane protein and in total protein (mNR2B/tNR2B) was calculated.Results Compared with group C,PWT was significantly decreased,PWL was shortened,the expression of mNR1,mNR2B,tNR1 and tNR2B was up-regulated,and the ratios of mNR1/tNR1 and mNR2B/tNR2B were increased in R + I,H1 and H2 groups.Compared with group R + I,PWT was significantly increased,PWL was prolonged,the expression of mNR1 and mNR2B was down-regulated,and the ratios of mNR1/tNR1 and mNR2B/tNR2B were decreased in Ht and H2 groups.Compared with group H1,PWT was significantly increased,PWL was prolonged,the expression of mNR1 and mNR2B was down-regulated,and the ratios of mNR1/tNR1 and mNR2B/tNR2B were decreased in group H2.Conclusion Hydrogen-rich saline can attenuate incisional pain-remifentanil-induced hyperalgesia in rats and inhibition of trafficking of spinal neuronal NR1 and NR2B from cytoplasm to cell membrane may be involved in the mechanism. Key words: Hydrogen; Hyperalgesia; Piperidines; Pain, postoperative

Effect of hydrogen-rich saline on neuropathic pain in rats

Objective To evaluate the effect of hydrogen-rich saline on neuropathic pain in rats.Methods One hundred and twenty male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 4 groups (n =30 each) using a random number table:sham operation group (S group); hydrogen-rich saline group (H group); chronic constrictive injury (CCI) group; CCI + hydrogen-rich saline group (CH group).Neuropathic pain was induced by CCI.The animals were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.The left sciatic nerve was exposed and 4 ligatures were placed on the sciatic nerve at 1 mm intervals.In H and CH groups,hydrogen-rich saline 10 ml/kg was injected intraperitoneally twice a day for 3 consecutive days starting from 1 day after CCI.The paw withdrawal threshold to mechanical stimulation with yon Frey filament (PWT) and paw withdrawal latency to thermal stimulation (PWL) were measured on the day before CCI (T0) and on 3,5 and 7 days after CCI (T1-3).The rats were sacrificed after the last measurement of pain threshold at T4.The left lumbar segments (L4-6) of the spinal cord were removed for determination of the contents of tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β),interleukin-6(IL-6) and high-mobility group box-1 (HMGB1) (by ELISA),and malondialdehyde (MDA) content and superoxide dismutase (SOD) and catalase (CAT) activities (using visible spectrophotometer).Results Compared with S group,PWT was significantly decreased,PWL was shortened,the contents of TNF-α,IL-1β,IL-6,HMGB1 and MDA were increased,and the activities of SOD and CAT were decreased at each time point after CCI in CCI and CH groups.Compared with CCI group,PWT was significantly increased,PWL was prolonged,the contents of TNF-α,IL-1β,IL-6,HMGB1 and MDA were decreased,and the activities of SOD and CAT were increased at all the time points after CCI in group CH.Conclusion Hydrogen-rich saline can alleviate neuropathic pain in rats via reducing the inflammatory response and lipid peroxidation response. Key words: Hydrogen; Neuralgia

Protective effect of hydrogen-rich saline on ischemia-reperfusion-induced liver injury in rats

Objective To study the effects of hydrogen-rich saline (HS) on the protection of hepatic injury induced by ischemia-reperfusion (I/R) in rats.Methods Male Sprague-Dawley rats (n =60) were randomly divided into three experimental groups:sham-operated group,I/R plus saline treatment group,and I/R plus hydrogen-rich saline treatment group.Reperfusion liver was conducted 180 mins after liver ischemia.Blood samples and liver tissues were collected.Result Serum ALT,AST levels,and MDA content,HMGB1,TNF-α,IL-1β and IL-6 levels in liver tissue were increased,but SOD and GSH activity were decreased significantly by I/R.Hydrogen-rich saline reduced oxidative stress and inflammatory reaction,and relieved morphological liver injury (P ＜ 0.01).Conclusion Hydrogen-rich saline attenuated I/R induced liver damage by reduction of oxidative injury and inflammatory reaction. Key words: Hydrogen-rich saline (HS) ; Ischemia-reperfusion (I/R) ; Oxidative injury

Effects of hydrogen-rich saline on early diabetic neuropathic pain in rats

Objective To evaluate the effects of hydrogen-rich saline on the early diabetic neuropathic pain (DPN) in rats.Methods Healthy male Sprague-Dawley rats,aged 8 weeks,weighing 180-220 g,were used in the study.DPN model was made by intraperitoneal injection of 1％ streptozocin (STZ) 65 mg/kg.Twelve diabetic rats were randomly divided into 2 groups (n =6 each) using a random number table:DPN group and hydrogen-rich saline group (HRS group).Another 6 normal rats were randomly collected as control group (group C).At 14 days after STZ injection,hydrogen-rich saline 5 ml/kg was injected intraperitoneally once a day for 14 consecutive days in HRS group,while C and DNP groups received the equal volume of normal saline.Mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and paw withdrawal latency to nociceptive thermal stimulation (TWL) were measured at 2 days before STZ injection (To) and 7,14,21 and 28 days after STZ injection (T1-4).The motor nerve conduction velocity (MNCV) of the right hindlimb and distal motor latency was measured after pain threshold was measured at T4.After measurement of neurological function was completed,tumor necrosis factor-α (TNF-α) and i nterleukin-6 (IL-6) contents (by ELISA) and nuclear factor kappa B (NF-κB) activity (by immuno-histochemistry) were detected.Results Compared with group C,MNCV was significantly decreased,the motor latency was prolonged,MWT was decreased at T1-T4,TWL was shortened at T2-T4,TNF-a and IL-6 contents were increased,and NF-κB activity was enhanced in DNP and HRS groups (P ＜ 0.05).Compared with group DNP,MNCV was significandy increased,the motor latency was shortened,MWT was increased at T3.4,TWL was prolonged at T4,TNF-a and IL-6 contents were decreased,and NF-κB activity was weakened in group HRS (P ＜ 0.05).Conclusion Hydrogen-rich saline can relieve the early DPN through inhibiting NF-κB signaling pathway in rats. Key words: Hydrogen; Diabetes mellitus; Neuralgia

Effect of hydrogen-rich saline on regulatory T cells in peripheral blood during global cerebral ischemia-reperfusion in rats

Objective To evaluate the effect of hydrogen-rich saline on the regulatory T cells （Tregs ） in the peripheral blood during global cerebral ischemia-reperfusion （I/R ） in rats .Methods Seventy-seven male Sprague-Dawley rats ,aged 2-3 months ,weighing 260-300 g ,were randomly divided into 3 groups using a random number table：sham operation group （group S , n=11） ,group I/R （n=33） ,and hydrogen-rich saline group （group H , n=33 ） .Global cerebral I/R was produced by 4-vessel occlusion method .The bilateral carotid arteries were blocked for 15 min followed by reperfusion in I/R and H groups .In group H ,0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected intraperitoneally at 0 and 6 h of reperfusion ,while the equal volume of normal saline was injected instead in the other two groups .Before ischemia （T0 ） in group S and at 6 ,24 and 72 h of reperfusion （T1-3 ） in I/R and H groups ,7 rats were chosen ,the blood samples from the peripheral vein were collected for determination of the number of Tregs . Then the animals were sacrificed and the spleen was removed for measurement of transforming growth factor-β1 （TGF-β1） content .The left 4 rats of each group were sacrificed at T0 and T1-3 and the brains were obtained for examination of the pyramidal cell morphology in the hippocampal CA 1 region and for determination of the number of pyramidal cells in brain tissues .Results Compared with group S , the number of pyramidal cells in the hippocampal CA1 region ,the number of Tregs in the peripheral blood and content of TGF-β1 in the spleen were significantly decreased at T1-3 in group I/R （ P〈0.05） .Compared with＆nbsp;group I/R ,the number of pyramidal cells in the hippocampal CA 1 region and the number of Tregs in the peripheral blood at T2-3 ,and the content of TGF-β1 in the spleen at T1-3 were significantly increased （ P〈0.05） ,and the pathological changes of pyramidal cells were attenuated in group H .Conclusion The mechanism by which hydrogen-rich saline attenuates global cerebral I/R injury may be related to the increased number of Tregs in peripheral blood and promoted secretion of TGF-β1 in rats . Key words: T-lymphocytes,regulatory; Hydrogen; Reperfusion injury; Brain

Protective effects of hydrogen-rich saline in uncontrolled hemorrhagic shock.

Hydrogen is considered to be a novel antioxidant as it inhibits inflammation, removes oxygen-derived free radicals and reduces oxidative damage. This study investigated the effects of hydrogen-rich saline on plasma interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD) and malondialdehyde (MDA) in rats with uncontrolled hemorrhagic shock (UHS). The UHS model was induced by arterial bleeding and tail amputation. The rats were randomly divided into: Group A (sham-operated group), Group B [shock + intravenously (IV) injected saline], Group C (shock + IV-injected hydrogen-rich saline), Group D [shock + intraperitoneally (IP) injected saline] and Group E (shock + IP-injected hydrogen-rich saline). The survival rate 24 h after successful resuscitation was calculated. The mean arterial pressure and heart rate were recorded at 0, 30, 90 and 210 min. The plasma levels of IL-6, TNF-α, SOD and MDA were measured at 0, 90 and 210 min. The survival rate of each group was 100% and the hemodynamics among the experimental groups were not significantly different. At 90 and 210 min, the levels of IL-6, TNF-α and MDA in Groups C and E were lower than those of Groups B and D, while the SOD levels were higher than those of Groups B and D (P<0.01). At 90 min, the levels of IL-6, TNF-α and MDA in Groups B and C were lower than those of Groups D and E, respectively (P<0.01). Hydrogen-rich saline has anti-inflammatory and anti-oxidative effects in UHS. In conclusion, the results showed that itravenous injection of hydrogen-rich saline is more effective than intraperitonal injection.

Protective Effects of Hydrogen-Rich Saline Against Erectile Dysfunction in a Streptozotocin Induced Diabetic Rat Model

Protective Effects of Hydrogen-Rich Saline Against Erectile Dysfunction in a Streptozotocin Induced Diabetic Rat Model

Protective effects of hydrogen-rich saline against γ-ray irradiation-induced injury in kidney epithelial HK-2 cells

Protective effects of hydrogen-rich saline against γ-ray irradiation-induced injury in kidney epithelial HK-2 cells

Long-term treatment of hydrogen-rich saline abates testicular oxidative stress induced by nicotine in mice

The present study was designed to test the hypothesis that long-term treatment with hydrogen-rich saline abated testicular oxidative stress induced by nicotine in mice. The effects of hydrogen-rich saline (6ml/kg, i.p.), vitamin C (60mg/kg, i.p.) and vitamin E (100mg/kg, i.p.) on reproductive system and testicular oxidative levels in nicotine-treated (4.5mg/kg, s.b.) mice were investigated. It was found that vitamin C and vitamin E attenuated serum oxidative level, but did not lower testicular oxidative levels in mice subjected to chronic nicotine treatment, and did not improve the male reproductive damage and apoptosis induced by nicotine. Different from normal antioxidants, vitamin C and vitamin E, hydrogen-rich saline abated oxidative stress in testis, and protected against nicotine-induced male reproductive damages. Our results first demonstrated that long-term treatment with hydrogen-rich saline attenuated testicular oxidative level and improved male reproductive function in nicotine-treated mice.

Protective effect of hydrogen on diabetic peripheral neuropathy by suppressing nulcear factor-kappaB pathway

Objective To investigate the effect of hydrogen on streptozotocin (STZ)-induced diabetic rat models with peripheral neuropathy and explore the possible mechanism. Methods Eighteen male Sprague⁃Dawley (SD) rats were randomly divided into model group (N = 6), hydrogen-treated group (N = 6) and normal control group (N = 6). After fasting for 12 h, experimental diabetic rat models were established by intraperitoneal injection of STZ (65 mg/kg of single dose), while normal control group only received a same dose of citrate buffer. Diabetes was confirmed with a fasting plasma glucose more than 16.67 mmol/L 48 h after STZ injection. After diabetic models were established successively, hydrogen-rich saline (5 ml/kg) was administered by intraperitoneal injection in hydrogen-treated group daily in 7th and 8th week after diabetes induction. Corresponding model and normal control groups received a same dose of normal saline. Changes of sciatic function and pain behavior in rats of different groups were measured to investigate the effect of hydrogen-rich saline. Proinflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and nuclear factor-kappaB (NF-κB) p65 expression were then determined to clarify the possible mechanism of hydrogen-mediated protection. Results 1) Compared with normal control group, body weight in model group decreased significantly, while plasma glucose levels increased significantly (P = 0.000, for all) 8 weeks after STZ induction. Hydrogen did not show any effects on body weight and plasma glucose levels of treated rat models in comparison with model group (P = 0.256, 0.821). 2) Compared with normal control group, motor nerve conduction velocity (MNCV), heat pain threshold (HPT) and mechanical withdrawal threshold (MWT) decreased significantly in model group (P = 0.000, for all), but increased significantly in hydrogen-treated group when compared with model group in the 8th week (P = 0.000, for all). 3) Hydrogen also reduced the positively expressed cells of NF-κB p65 (P = 0.000) as well as levels of TNF-α and IL-6 (P = 0.000, for all). Conclusion Inflammation may participate and exaggerate painful diabetic neuropathy. Besides, hydrogen has the protective potential of ameliorating neuroinflammation and peripheral nerve injury by suppressing NF-κB pathway and its downstream inflammatory cytokines.

Protective effects of hydrogen-rich saline on acute kidney injury in rats with sepsis

Objective To investigate the effects of hydrogen-rich saline (HRS) on acute kidney injury (AKI) in a rat model of sepsis.Methods Twenty-four male SD rats were randomly (random number) assigned to the sham operation group,the cecal ligation and puncture (CLP) group,and the CLP + HRS group (n =8).The rats in the sham operation group were treated with lapa(r ly) without CLP,while the rats in the CLP group and CLP + HRS group were made to be CLP-induced sepsis model.At 0,6 and 18 h after sham operation or CLP,rats in the CLP + HRS group received an intraperitoneal injection of HRS of 5 ml/kg,and 5 ml/kg of normal saline was given to the rats in the sham group and CLP group.Plasma levels of TNF-α,IL-6,IL-10,and HMGB1 were determined by enzyme linked immunosorbent assay (ELISA),plasma BUN and Cr were assayed by enzymatic assays,renal malondialdehyde (MDA) levels and superoxide dismutase (SOD) activities were determined by xanthine oxide method,renal tissue injury was dertermined by hematoxylin-eosin staining (liE),and renal cell apoptosis was assessed by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) at 24 h after sham operation or CLP.Statistical analysis carried out by using one way analysis of variance followed by Student-Newman-Keuls (SNK) test.Results Compared with the sham group,rats in the CLP and CLP + HRS groups had increases in plasma levels of TNF-α,IL-6,IL-10,HMGB1,BUN,Cr,renal MDA,severe renal injury and apoptosis,and a decrease in renal SOD activities (P ＜ 0.05).Compared with the CLP group,the CLP + HRS group had decreases in plasma levels of HMGB1,BUN,renal MDA,mild renal injury and apoptosis,and an increase in renal SOD activities (P ＜ 0.05).Conclusions HRS reduces inflammatory reaction and oxidative stress,improving renal function during AKI in the rat model of sepsis. Key words: Hydrogen-rich saline ; Sepsis ; Inflammatory reaction ; Oxidative stress ; Acute kidney injury

Effect of hydrogen-rich saline on allergic contact dermatitis in mice

Objective To evaluate the therapeutic effect of hydrogen-rich saline on allergic contact dermatitis (ACD) in mice and to explore its underlying mechanisms.Methods Forty mice were equally divided into 4 groups:control group,control treatment group,ACD group and ACD treatment group.ACD was induced by repetitive topical application of dinitrofluorobene (DNFB) to the left ear of mice on day 1,2 and 5.Hydrogen-rich saline was intraperitoneally given to the mice in the ACD treatment group at a dose of 5 ml/kg per day from day 1 to 5.On day 6,the mice were sacrificed,ear tissue was removed from them and subjected to measurement of thickness and weight,detection of tumor necrosis factor (TNF)-α,interleukin (IL)-6,IL-17 and interferon (IFN)-γ expression by enzyme linked immunosorbent assay,as well as numeration of inflammatory cells in lesions after hematoxylin-eosin (HE) staining.Data were processed by SPSS 18.0 software,and statistical analysis was carried out by one-way analysis of variance and least significant difference (LSD) procedure.Results Compared with the control group,the ACD group showed a significant increase in lesion score (7.33 ± 1.53 vs.0,P ＜ 0.05),differences in the thickness ((0.73 ± 0.15) mm vs.(0.13 ± 0.05) mm,P ＜ 0.05) and swelling degree (expressed as tissue weight:(18.67 ± 3.05) mg vs.(3.33 ± 1.52) mg,P ＜ 0.05) between the left and right ear,expressions of TNF-α ((1475.52 ± 233.81) pg/mg vs.(239.01 ± 52.39) pg/mg,P＜ 0.05),IL-6 ((184.65 ± 78.39) pg/mg vs.(42.28 ± 17.64) pg/mg,P＜ 0.05),IL-17 ((628.56 ± 201.44) pg/mg vs.(127.58 ± 50.28) pg/mg,P＜ 0.05) and IFN-γ ((197.72 ± 37.81) pg/mg vs.(24.57 ± 8.31) pg/mg,P ＜ 0.05),and the number of inflammatory cells per square millimetre in the left ear tissue (752.00 ± 166.06 vs.127.33 ± 77.18,P ＜ 0.05).However,hydrogen-rich saline treatment induced a statistical decrease in all of these parameters in the ACD treatment group compared with the ACD group,including lesion score (3.33 ± 0.58,P ＜ 0.05),difference in thickness ((0.46 ± 0.11) mm,P ＜ 0.05) and swelling degree ((11.00 ± 2.64) mg,P ＜ 0.05),expressions of TNF-α ((817.72 ± 101.13) pg/mg,P＜ 0.05),IL-6 ((95.86 ± 36.65) pg/mg,P＜ 0.05),IL-17 ((373.38 ± 126.74) pg/mg,P＜ 0.05),IFN-γ ((63.31± 17.38) pg/mg,P ＜ 0.05) and the number of inflammatory cells per square millimetre (384.00 ± 97.35,P ＜0.05).Conclusion Hydrogen may inhibit the release of inflammatory factors in ACD.

Effect of hydrogen-rich saline on TNF-α, NF-κB and apoptosis in skin flap after ischemia/reperfusion injury

Objective To study the protective effect of hydrogen-rich saline (HRS) on apoptosis in skin flap after ischemia/reperfusion injury.Methods Total 18 Sprague-Dawley rats were randomly divided into three groups:a HRS treated group and two physiological saline treated groups (controls 1,2).The rats were anesthetized and an extended abdominal skin flap (6 cm × 9 cm) was elevated in each animal.Ischemia was induced by clamping the left right pedicle for 3 h,then HRS was administered intraperitoneally 10 min before reperfusion,and physiological saline was injected in control groups 1 and 2.In the control group 2,the flaps were elevated without occluding the artery and vein.Five days postoperation,apoptosis,TNF-α level in flap were measured with ELISA,NF-κB in nucleus was determined by Westernblot.Results Apoptotic rate represented (39.72±8.09) ％in HRS group and (69.43±13.27) ％ in control group 1,respectively.Treatment with HRS resulted in a marked reduction in apoptotic rate.TNF-α level was (516.408±38.674) pg/ml in the control group 1,a significant reduced TNF-α was measured in HRS group,accounting for (269.136 ±24.530) pg/ml.Moreover,NF-κB activation was significantly down-regulated by HRS.In control group 2,no significant apoptosis was observed because of non-blood occlusion,and there was no marked elevation of TNF-α and NF-κB.Conclusions HRS can protect skin from ischemia/reperfusion injury,attenuate apoptosis in flaps,which may be associated with the inhibition of TNF-α and NF-kb elevation. Key words: Hydrogen-rich saline(HRS); Skin flap; Apoptosis; TNF-κB; TNF-α

Effects of Hydrogen-Rich Saline on Taurocholate-Induced Acute Pancreatitis in Rat

Oxidative stress plays an important role in the pathogenesis of acute pancreatitis (AP). As an ideal exterminator of poisonous free radicals, hydrogen can clearly reduce the degree of oxidative damage caused by severe acute pancreatitis (SAP) and lessen the presence of inflammatory cytokines. The aim of this study was to investigate the effects and mechanism of hydrogen-rich saline on SAP in rats. Serum TNF-α, IL-6, and IL-18 and histopathological score in the pancreas were reduced after hydrogen-rich saline treatment. Malondialdehyde (MDA) and myeloperoxidase (MPO) contents were obviously reduced, while superoxide dismutase (SOD) and glutathione (GSH) contents were increased after hydrogen-rich saline treatment. The expression of mRNA of tumor necrosis factor-α (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in the pancreas was reduced in hydrogen-rich saline treated group. In conclusion, intravenous hydrogen-rich saline injections could attenuate the severity of AP, probably via inhibiting the oxidative stress and reducing the presence of inflammatory mediators.

Hydrogen-Rich Saline Prevents Early Neurovascular Dysfunction Resulting from Inhibition of Oxidative Stress in STZ-Diabetic Rats

ABSTRACTPurpose: Diabetic retinopathy (DR) is characterized by increased oxidative and nitrosative stress, both of which lead to neurotoxicity and vascular permeability. Previous studies on a variety of organs indicate that hydrogen-rich saline not only has considerable antioxidant and anti-inflammatory properties, but also suppresses oxidative stress-induced injury. In the present study, we assessed the effects of hydrogen-rich saline on neurovascular dysfunction and oxidative stress in an animal model (rat) of DR.Materials and Methods: Male Sprague-Dawley rats with streptozotocin (STZ)-induced diabetes mellitus (DM) were injected intraperitoneally with 5 ml/kg hydrogen-saturated (experimental) or plain (control) saline daily for one month. Visual function and blood–retinal barrier (BRB) integrity were evaluated by electroretinography (ERG) and bovine serum albumin (BSA)-fluorescence, respectively. Histological changes in the inner retina were assessed by light microscopy. Biomarkers of oxidative stress, including 4-hydroxynonenal (4-HNE) and 8-hydroxy-2-deoxyguanosine (8-OH-dG), and antioxidant enzymes, including superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione transferase, were evaluated by ELISA. Synaptophysin and brain-derived neurotrophic factor (BDNF) levels were measured by immunoblotting.Results: STZ-diabetic rats were marked by clearly reduced b-wave amplitudes and oscillatory potentials, DM-related BRB breakdown and histological changes in the inner retina, all of which were suppressed following treatment with hydrogen-rich saline. Furthermore, hydrogen-rich saline reduced oxidative stress, increased antioxidant enzyme activities and preserved synaptophysin and BDNF levels in the diabetic rat retina.Conclusions: Based on its inhibition of oxidative stress and up-regulation of anti-oxidative enzymes, we conclude that hydrogen-rich saline is a potentially valuable therapeutic modality for the treatment of DR.

Effects of hydrogen-rich saline on postoperative cognitive dysfunction in aged mice

Objective To investigate the effects of hydrogen-rich saline on postoperative cognitive dysfunction (POCD) in aged mice and the possible mechanism.Methods Seventy-two male C57BL/6 mice,aged 18 months,weighing 45-55 g,were randomly divided into 4 groups (n =18 each):control group (group C),hydrogen-rich saline group (group H),POCD group and POCD + hydrogen-rich saline group (group PH).POCD was induced by open tibial fracture.Hydrogen-rich saline 10 ml/kg was injected intraperitoneally twice a day for 7 days before surgery.Six mice were chosen at day 1,3 and 7 after surgery and cognitive function was assessed by Morris water maze test.The mice were then sacrificed and hippocampus was isolated for determination of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) contents (by ELISA) and nuclear factor-κB (NF-κB) activity.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased at 3 and 7 days after surgery,and the contents of NF-α and IL-1β and NF-κB activity were increased during 1,3 and 7 days after surgery in group POCD (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group H (P ＞ 0.05).Compared with group POCD,the escape latency was significantly shortened,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased at 3 and 7 days after surgery,and the contents of NF-α and IL-1β and NF-κB activity were decreased at 1,3 and 7 days after surgery.Conclusion Hydrogen-rich saline can alleviate POCD via inhibiting NK-κB activity in hippocampus and reducing inflammatory response. Key words: Hydrogen ; Cognition disorders ; NF-kappa B

Experimental studies of effects of hydrogen-rich saline in rats with severe acute pancreatitis

To explore the mechanism and effectiveness of inducing severe acute pancreatitis (SAP) with saline saturated hydrogen in rats. Based on a random number table, 54 Sprague-Dawley rats were divided into 3 groups: control, SAP and treatment (n = 18 each). The model of SAP was established by a retrograde injection of 5% sodium taurocholate into biliary-pancreatic duct. The treatment group received an injection of 5 ml/kg hydrogen-rich saline into tail vein at 1 h post-modeling. The control group underwent only pancreatic tipping. All rats were exsanguinated under anesthesia by aortal puncture after 24 hours. The serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-18 (IL-18) in 3 groups were measured by enzyme-linked immunosorbent assay (ELISA). The expression of TNF-αmRNA in pancreatic tissue was measured by reverse transcription (RT)-PCR. And intercellular adhesion molecule-1 (ICAM-1) in pancreatic tissue was measured by immunohistochemistry. The pancreatic tissues were harvested to examine the microscopic changes. The levels of malonic aldehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and glutathione (GSH) in pancreatic tissue was measured by specific kits. The pathological scores of pancreatic tissue were significantly lower in the treatment group than those in the SAP group (9.7 ± 2.0 vs 13.2 ± 2.5, P < 0.05). The serum level of TNF-αwas significantly lower in the treatment group than that in the SAP group ((88 ± 17) vs (1171 ± 18) pg/ml, P < 0.05). And the serum levels of IL-6 and IL-18 were lower in the treatment group than those in the SAP group ((25.2 ± 5.9) vs (37.3 ± 4.6) ng/L, (401 ± 100) vs (439 ± 103) ng/L, both P < 0.05). As compared with the SAP group, the levels of MDA and MPO decreased significantly in the treatment group ((8.2 ± 2.8) vs (14.7 ± 2.7) nmol/mg, (0.040 ± 0.011) vs (0.170 ± 0.071) U/g, both P < 0.05). However, the levels of SOD and GSH significantly higher than those of the treatment group ((22.1 ± 1.3) vs (15.1 ± 1.7) U/mg, (4.8 ± 0.4) vs (2.5 ± 0.3) U/mg, both P < 0.05). The expressions of TNF-α mRNA and ICAM-1 in pancreatic tissues were lower than those of the treatment group (0.33 ± 0.17 vs 0.94 ± 0.31, 2.0 ± 0.4 vs 2.8 ± 0.8, both P < 0.05). Hydrogen-rich saline can decrease the levels of inflammatory mediators and reduce the pathological damage of pancreas through the inhibition of oxidative stress.

Effects of Hydrogen-rich Saline on Rats with Acute Carbon Monoxide Poisoning

BackgroundStudies have shown that inhalation of hydrogen gas, which acts as an antioxidant, can protect the brain against free radicals in rats with ischemia-reperfusion. The neuronal damage caused by acute carbon monoxide (CO) poisoning is partly free radical mediated. We hypothesize that hydrogen may prevent neurological damage from CO poisoning. ObjectivesThis study is designed to test whether hydrogen (H2)-rich saline will have a protective effect on rats with acute CO poisoning. MethodsMale Sprague-Dawley rats were subjected to CO poisoning. H2-rich saline was administered by peritoneal injection (6mL/kg/24h). We used the Morris water maze and the open field test to determine cognitive function. After cognitive function studies, rats were decapitated and the levels of trace elements copper (Cu), zinc (Zn), and iron (Fe) in serum and brain were assessed by flame atomic absorption spectrometry. Necrosis, apoptosis, and autophagy of neurons were assessed by H-E staining and immunohistochemical staining in another group of rats. ResultsH2-rich saline treatment improved the cognitive deficits and reduced the degree of necrosis, apoptosis, and cell autophagy in rats. Additionally, H2-rich saline decreased the content of Fe in serum and brain in these rats, and increased the content of serum Cu related to free radical metabolism. ConclusionsH2-rich saline may effectively protect the brain from injury after acute CO poisoning. The mechanism of this protection may be related to lessening oxidative damage by affecting trace elements in vivo.

Effects of hydrogen-rich saline given during reperfusion on global cerebral ischemia-reperfusion injury in rats

Objective To evaluate the effect of hydrogen-rich saline given during reperfusion on global cerebral ischeraia-reperfusion (I/R) injury in rats.Methods Seventy-two adult male SD rats,aged 2.0-2.5 months,weighing 260-300 g,were randomly divided into 3 groups (n ＝ 24 each):sham operation group (group S),group I/R and hydrogen-rich saline group (group H).In groups I/R and H cerebral I/R was induced by occlusion of 4 vessels( cauterization of bilateral carotid arteries and 15 min occlusion of bilateral common carotid arteries).In group H intraperitoneal 0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected at 6 h of reperfusion,while equal volume of normal saline was injected instead of hydrogen-rich saline.Eighteen rats of each group were sacririced at 24 h of reperfusion,and then the hippocampi were removed for determination of malondialdehyde (MDA),tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6)contents,and nuclear factor-κB (NF-κB) activity and activated caspase-3 expression.Another six rats of each group were sacrificed at 72 h of reperfusion,and then brain tissues were removed for microscopic examination and counting the number of uninjured pyramidal cells in hippocampal CA1 region.Results Compared with S group,the contents of MDA,TNF-α,IL-6 and NF-κB activity were significantly increased,activated caspase-3 expression was significantly up-regulated,uninjured pyramidal cells in hippocampal CA1 region were significantly decreased in I/R group( P < 0.05).Hydrogen-rich saline given during reperfusion attenuated the above-mentioned I/R-induced changes( P < 0.05 ).The histologic damage of the hippocampal CA1 region was significantly slighter in group H than group I/R.Conclusion Hydrogen-rich saline given during reperfusion can reduce global cerebral I/R injury in rats through inhibition of lipid peroxidation,inflammatory response and apoptosis. Key words: Hydrogen; Reperfusion injury; Brain

Protective effects of hydrogen-rich saline against hind limb ischemia reperfusion injury and associated remote lung injury

Objective To explore the protective effects of hydrogen-rich saline against hind limb ischemia reperfusion injury and associated remote lung injury in rats. Methods Thirty Sprague-Dawley rats were randomly divided into three groups: sham group ( group S) ; saline control group (group C) , undergoing intravenous infusion of saline (1 ml/100 g) at 10 min before reperfusion; hydrogen-treated group ( group H) , undergoing intravenous infusion of hydrogen-rich saline (1 ml/100 g) at 10 min before reperfusion. Hind limb ischemia reperfusion model was established by 4 h of the abdominal aorta ligation followed by a 4 h of reperfusion. Rats were killed at the end of reperfusion, and blood samples were collected for determination of serum levels of tumor necrosis factor-α ( TNF-α) , interleukin-6 (IL-6) and malonyldialdehyde ( MDA). Specimens of lung and gastrocnemius muscle were collected for measurement of wet-todry weight (W/D) ratio, light microscopic examination and detection of plasma TNF-α, IL-6 and MDA.Results The pathological changes were milder in group H than in group C. The lung (4. 47 ±0. 23) and gastrocnemius muscle (4. 04 ±0. 22) W/D ratio in group H was significantly lower than in group C (4. 67 ±0.27 vs4. 67 ±0.27, P＜0.05). The serum [(2.08 ± 1. 27) μmol/L, (528. 22 ±221. 17) , (41.14± 13. 01) ng/L, respectively] and homogenate levels [(3. 58 ± 1. 62) μmol/L, ( 1. 17 ±0. 15) , (0. 79±0.29) ng/L in the lung; (8.91 ±3. 13) μmol/L, (3. 13 ±0. 41) ,(3. 07 ± 1. 69) ng/L in the gastrocnemius muscle, respectively] of MDA, IL-6 and TNF-a were also significantly lower in group H than in group C (P ＜0. 05) , but there was no significant difference from group S. Conclusion Infusion of hydrogen-rich saline before reperfsion significantly alleviates the hind limb ischemia reperfusion injury and associated remote lung injury. Key words: Hydrogen-rich saline; Lung; Skeletal muscle; Reperfusion injury; Free radicals