Jianmei Cai

Neuroprotective effects of hydrogen saline in neonatal hypoxia-ischemia rat model.

Cerebral hypoxia-ischemia (HI) represents a major cause of brain damage in the term newborn. This study aimed to examine the short and long-term neuroprotective effect of hydrogen saline (H(2) saline) using an established neonatal HI rat pup model. Seven-day-old rat pups were subjected to left common carotid artery ligation and then 90 min hypoxia (8% oxygen at 37 degrees C). H(2) saturated saline was administered by peritoneal injection (5 ml/kg) immediately and again at 8 h after HI insult. At 24 h after HI, the pups were decapitated and brain morphological injury was assessed by 2,3,5-triphenyltetrazolium chloride (TTC), Nissl, and TUNEL staining. Acute cell death, inflammation and oxidative stress were evaluated at 24 h by studying caspase-3 activity, MDA measurement as well as Iba-1 immunochemistry in the brain. At 5 weeks after HI, spontaneous activity test and Morris water maze test were conducted. We observed that H(2) saline treatment reduced the caspase activity, MDA, Iba-1 levels, the infarct ratio, and improved the long-term neurological and neurobehavioral functions. H(2) saline has potentials in the clinical treatment of HI and other ischemia-related cerebral diseases.

Hydrogen therapy reduces apoptosis in neonatal hypoxia-ischemia rat model.

Hypoxia-ischemia (HI) brain injury is a major cause of neuronal cell death especially apoptosis in the perinatal period. This study was designated to examine the effect of hydrogen therapy on apoptosis in an established neonatal HI rat pup model. Seven-day-old rat pups were subjected to left common carotid artery ligation and then 90 min hypoxia (8% oxygen at 37 degrees C). Immediately after HI insult, pups were placed into a chamber filled with 2% H2 for 30 min, 60 min, or 120 min, respectively. 24 h after 2% H2 therapy, the pups were decapitated and brain injury was assessed by 2,3,5-triphenyltetrazoliumchloride (TTC), Nissl, and TUNEL staining, as well as caspase-3, caspase-12 activities in the cortex and hippocampus. H2 treatment in a duration-dependent manner significantly reduced the number of positive TUNEL cells and suppressed caspase-3 and -12 activities. These results indicated H2 administration after HI appeared to provide brain protection via inhibition of neuronal apoptosis.

Hydrogen-rich saline protects against intestinal ischemia/reperfusion injury in rats

Hydrogen gas was reported to reduce reactive oxygen species and alleviate cerebral, myocardial and hepatic ischemia/reperfusion (I/R) injuries. This paper studied the effect of hydrogen-rich saline, which was easier for clinical application, on the intestinal I/R injury. Model of intestinal I/R injury was induced in male Sprague-Dawley rats. Physiological saline, hydrogen-rich saline or nitrogen-rich saline (5 ml/kg) was administered via intravenous infusion at 10 min before reperfusion, respectively. The intestine damage was detected microscopically and was assessed by Chiu score system after I/R injury. In addition, serum DAO activity, TNF-α, IL-1β and IL-6 levels, tissue MDA, protein carbonyl and MPO activity were all increased significantly by I/R injury. Hydrogen-rich saline reduced these markers and relieved morphological intestinal injury, while no significant reduction was observed in the nitrogen-rich saline-treated animals. In conclusion, hydrogen-rich saline protected the small intestine against I/R injury, possibly by reduction of inflammation and oxidative stress.

Hydrogen-rich saline protects myocardium against ischemia/reperfusion injury in rats.

Protective effect of hydrogen (H2) gas on cardiac ischemia-reperfusion (I/R) injury has been demonstrated previously. This study was designed to test the hypothesis that hydrogen-rich saline (saline saturated with molecular hydrogen), which is easy to use, induces cardioprotection against ischemia (30 min) and reperfusion (24 h) injury in rats. Adult male Sprague-Dawley rats underwent 30-min occlusion of the left anterior descending (LAD) coronary artery and 24-h reperfusion. Intraperitoneal injection of hydrogen-rich saline before reperfusion significantly decreased plasma and myocardium malondialdehyde (MDA) concentration, decreased cardiac cell apoptosis, and myocardial 8-hydroxydeoxyguanosine (8-OHdG) in area at risk zones (AAR), suppressed the activity of caspase-3, and reduced infarct size. The heart function parameters including left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVDP), +(dP/dt)max and −(dP/dt)max were also significantly improved 24 h after reperfusion. It is concluded that hydrogen-rich saline is a novel, simple, safe, and effective method to attenuate myocardial I/R injury.

Hydrogen-rich saline reduces lung injury induced by intestinal ischemia/reperfusion in rats.

OBJECTIVE Hydrogen has been reported to selectively reduce the hydroxyl radical, the most cytotoxic of reactive oxygen species. In this study we investigated the effects of hydrogen-rich saline on the prevention of lung injury induced by intestinal ischemia/reperfusion (I/R) in rats. METHODS Male Sprague-Dawley rats (n=30, 200-220g) were divided randomly into three experimental groups: sham operated, intestinal I/R plus saline treatment (5ml/kg, i.v.), and intestinal I/R plus hydrogen-rich saline treatment (5ml/kg, i.v.) groups. Intestinal I/R was produced by 90min of intestinal ischemia followed by a 4h of reperfusion. RESULTS Hydrogen-rich saline treatment decreased the neutrophil infiltration, the lipid membrane peroxidation, NF-kappaB activation and the pro-inflammatory cytokine interleukin IL-1beta and TNF-alpha in the lung tissues compared with those in saline-treated rat. CONCLUSION Hydrogen-rich saline attenuates lung injury induced by intestinal I/R.

Hydrogen-rich saline improves memory function in a rat model of amyloid-beta-induced Alzheimer's disease by reduction of oxidative stress.

This study is to examine if hydrogen-rich saline reduced amyloid beta (Abeta) induced neural inflammation, and learning and memory deficits in a rat model. S-D male rats (n=84, 280-330g) were divided into three groups, sham-operated, Abeta1-42 injected and Abeta1-42 plus hydrogen-rich saline-treated animals. Hydrogen-rich saline (5ml/kg, i.p., daily) was injected for 14days after intracerebroventricular injection of Abeta1-42. The levels of MDA, IL-6 and TNF-alpha were assessed by biochemical and ELISA analysis. Morris Water Maze and open field task were used to assess the memory dysfunction and motor dysfunction, respectively. LTP were used to detect the electrophysiology changes, HNE and GFAP immunohistochemistry were used to assess the oxidative stress and glial cell activation. After Abeta1-42 injection, the levels of MDA, IL-6, and TNF-alpha were increased in brain tissues and hydrogen-rich saline treatment suppressed MDA, IL-6, and TNF-alpha concentration. Hydrogen-rich saline treatment improved Morris Water Maze and enhanced LTP in hippocampus blocked by Abeta1-42. Furthermore, hydrogen-rich saline treatment also decreased the immunoreactivitiy of HNE and GFAP in hippocampus induced by Abeta1-42. In conclusion, hydrogen-rich saline prevented Abeta-induced neuroinflammation and oxidative stress, which may contribute to the improvement of memory dysfunction in this rat model.

Sulforaphane protects brains against hypoxic-ischemic injury through induction of Nrf2-dependent phase 2 enzyme.

Neonatal hypoxia-ischemia (HI) brain injury involves reactive oxygen species (ROS) and inflammatory responses. Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, has cytoprotective effects against oxidative stress and its effect was mediated by NF-E2-related factor-2 (Nrf2), a transcription factor, and heme oxygenase 1 (HO-1) which is one of Nrf2 downstream target genes. This study was undertaken to investigate the neuroprotective mechanisms of SFN in a neonatal HI rat model. Seven-day-old rat pups were subjected to left common carotid artery ligation and hypoxia (8% oxygen at 37 degrees C) for 90 min. SFN (5mg/kg) was systemically administered 30 min before HI insult. Brain injury was assessed by 2,3,5-triphenyltetrazoliumchloride (TTC), Nissl, TUNEL staining, malondialdehyde (MDA), 8OH-dG level, and caspase-3 activity in the cortex and hippocampus. SFN pretreatment increased the expression of Nrf2 and HO-1 in the brain and reduced infarct ratio at 24h after HI. The number of TUNEL-positive neurons as well as activated macroglia and the amount of 8OH-dG, were markedly reduced after SFN treatment, accompanied by suppressed caspase-3 activity and reduced lipid peroxidation (MDA) level. These results demonstrated that SFN could exert neuroprotective effects through increasing Nrf2 and HO-1 expression.

Hydrogen-Rich Saline Provides Protection Against Hyperoxic Lung Injury

BACKGROUND Hydrogen has been proven to be a novel antioxidant through its selectively reducing of the hydroxyl radical. In this study, we investigated the effects of hydrogen-rich saline on the prevention of acute lung injury induced by hyperoxia (HALI) in rats. MATERIALS AND METHODS Physiologic saline, hydrogen-rich saline, or nitrogen-rich saline was administered through intraperitoneal (i.p.) injection during exposure to hyperoxia (10 mL/Kg), respectively. RESULTS Severity of HALI was assessed by the volume of pleural effusion, wet-to-dry weight ratio (W/D), and histologic analysis. Apoptosis in lung cells was determined with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive staining. The content of pro-inflammatory cytokine interleukin IL-1b and TNF-a in the lung tissues were detected by enzyme-linked immunosorbent assay (ELISA). Hydrogen-rich saline treatment provides protection against HALI by inhibiting lipid, DNA oxidation, and tissue edema. Moreover, hydrogen-rich saline treatment could inhibit apoptosis and inflammation while no significant reduction was observed in nitrogen-rich saline treated animals. CONCLUSION The results of this study demonstrate that hydrogen-rich saline ameliorated hyperoxia-induced acute lung injury by reducing oxidative stress and inflammatory cascades in lung tissue.

Protective Effects of Hydrogen Saline on Diabetic Retinopathy in a Streptozotocin-Induced Diabetic Rat Model

PURPOSE Diabetic retinopathy is the leading cause of blindness in the working population of the developed countries and also a significant cause of blindness in the elderly. This study aimed at examining the protective effect of H(2) saline on diabetic retinopathy in a streptozotocin-induced diabetic rat model. METHODS Sprague-Dawley male rats were divided into 3 groups as follows: (1) nondiabetic control group (non-DM control); (2) diabetic control group (DM control); and (3) diabetic rats receiving H(2) saline therapy (DM H(2) saline). Rats in DM H(2) saline group were intraperitoneally injected with H(2) saturated saline (5 mL/kg) every day for 4 weeks. Retinal vascular permeability was assessed by measuring Evans blue leakage into the retina. Retinal apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining and measuring caspase-3 activity. Retinal thickness was observed by hematoxylin and eosin staining. RESULTS Our results showed that H(2) saline treatment could depress the caspase activity, reduce the retinal apoptosis, and vascular permeability. The H(2) saline could also prominently attenuate the retinal parenchyma thickening that resulted from diabetic retinopathy. CONCLUSIONS Our preliminary studies indicated that H(2) saline may have potentials in the clinical treatment of diabetic retinopathy.

Protective effects of carboxyfullerene in irradiated cells and BALB/c mice

Abstract It has been reported that carboxyfullerene (C3) exhibits a strong free radical scavenging capacity, which plays a critical role in radiation damage. We hypothesized that C3 is an effective radioprotective agent. In this study, we demonstrated that C3 effectively scavenged hydroxyl radical in a cell-free Fenton system and that C3 showed no obvious toxicity for cultured cells. Different concentrations (100–400 mg/L) of C3 pretreatment effectively protected AHH-1 cells from radiation-induced apoptosis, by about 2-folds, while only 100 mg/L of C3 reduced the apoptosis rate of human intestinal crypt epithelial cells (HIECs). C3 also alleviated DNA damage detected using comet assay at 0, 4, and 8 h after irradiation. It was found that 75% of mice were protected from 7.2 Gy γ-irradiation-induced death when 100 mg/kg of C3 was administered prior to irradiation, but no change was observed for the survival time of mice which died. We also found that C3 attenuated radiation-mediated decreases in endogenous antioxidants such as SOD and GSH and reduced the level of MDA. In conclusion, these data showed that C3 effectively protected cells and mice from radiation injury, thus indicating the potential of C3 as a safe and effective radioprotectant.