Rei Edamatsu

Increased SOD activities and decreased lipid peroxide levels induced by low dose X irradiation in rat organs

After a single exposure to doses ranging from 0.05 to 0.50 Gy, the SOD activities in immune organs of the irradiated rats, at 4 h after whole-body X irradiation, showed significant increases by 50 to 90% (hereinafter p less than 0.05) in comparison with the control groups. The dose in the brain and liver showed tendencies to increase by 30 to 50% for doses ranging from 0.50 to 2.50 Gy. The levels of TBARS (lipid peroxides) in immune organs decreased significantly by 20 to 50% in contrast with the control groups for doses ranging from 0.10 to 1.00 Gy. Decreases by 20 to 30% were observed in brains and livers for doses near 0.50 Gy. When the dose was 0.25 Gy, the SOD activities in the spleen showed a persistent radiation-induced increase for at least 12 weeks, livers for 8 weeks, brains and thymuses for 4 weeks, and bone marrows for about 1 week. The TBARS levels in the brain and thymus showed persistent decreases due to irradiation for at least 12 weeks, and those in bone marrows for 8 h.

Increased Superoxide Dismutase Activity in Aged Human Cerebrospinal Fluid and Rat Brain Determined by Electron Spin Resonance Spectrometry Using the Spin Trap Method

Abstract: Superoxide dismutase (SOD) activity in CSF of patients was determined by electron spin resonance spec‐trometry using the spin trap method. Variation in SOD activity was found among patients. SOD activity in CSF of subjects increased with age and this was identified as Cu,Zn‐SOD activity by electrophoresis. In addition, animal experiments showed that SOD activities were higher in mitochon‐drial and cytosol fractions of aged rats than in those of adult rats. This finding on aged rat brain validates the increase of SOD activity in aged human CSF.

Antioxidant action of guilingji in the brain of rats with FeCl3-induced epilepsy.

The effects of Guilingji, an antiaging prescription comprised of traditional Chinese medicinal herbs and animal components, on the brain level of thiobarbituric acid reactive substances (TBARS) and superoxide dismutase (SOD) activity in the brain of rats with FeCl3-induced epilepsy were examined with fluorophotometry and electron spin resonance (ESR) spectrometry, respectively. The results showed that pretreatment of rats with Guilingji decreased the levels of TBARS in the left cortex, midbrain, and olfactory lobe, and increased the SOD activity in the midbrain and hypothalamus. These results are the first to demonstrate that Guilingji may possess an antiepileptic effect further to its proposed antiaging action, because decreasing the TBARS elevation and increasing the SOD attenuation in the brain are suggested to be important characteristics of antiepileptic agents. These results suggest that both the proposed antiaging and antiepileptic effects of Guilingji may operate through the mechanism of antioxidation.

Effect of hyperbaric oxygenation on the Na+, K+-ATPase and membrane fluidity of cerebrocortical membranes after experimental subarachnoid hemorrhage

It is reported that CNS hemorrage causes membrane dysfunction and may exacerbate this damage as a result of secondary ischemia or hypoxia. Since hyperbaric oxygenation improves oxygen metabolism, it may reduce this membrane damage. The present study was conducted to reveal whether hyperbaric oxygenation influences membrane alteration after hemorrhage. Thirty minutes after subarachnoid hemorrhage induction, rats were treated with hyperbaric oxygenation 2 ATA for 1 hour. Rats were decapitated 2 hours after subarachnoid hemorrhage induction. Na+, K+-ATPase activity measurement, and spin-label studies were performed on crude synpatosomal membranes. Subarachnoid hemorrhage decreased Na+, K+-ATPase activity. Spin label studies showed that hydrophobic portions of near the membrane surface became more rigid and the mobility of the membrane protein labeled sulfhydryl groups decreased after subarachnoid hemorrhage. Hyperbaric oxygenation significantly ameliorated most of the subarachnoid hemorrhage induced alterations. We conclude that hyperbaric oxygenation may be a beneficial treatment for acute subarachnoid hemorrhage.

Biochemical pathogenesis of post-traumatic epilepsy

Head trauma is often followed by epilepsy and may be related to the breakdown of red blood cells and hemoglobin within the CNS. Injection of hemoglobin or iron salts into the rat cortex is known to induce a chronic epileptic focus. We observed the formation of superoxide anion (O2) and hydroxyl radical (·OH) after ferric chloride injection into the rat cerebral cortex and suggest that these radicals, especially ·OH, may be responsible for the initiation of lipid peroxidation in neuronal membranes and for the accelerated production of guanidine compounds in the brain, which may in turn lead to epileptogenicity. Then, we found that treatment with epigallocatechin (EGC) or a phosphate diester of vitamins E and C (EPC), which are potent ·OH scavengers, significantly inhibited the formation of malondialdehyde and epileptic discharges in the iron-induced epileptic focus.

Effects of low-dose X-ray irradiation on biomembrane in brain cortex of aged rats.

We previously found that low-dose X-ray irradiation or radon (weak alpha-ray) inhalation increases SOD activities and reduces lipid peroxide levels in various organs of 7-week-old rats or rabbits. In this study, we examined how the changes of SOD activity, lipid peroxide level, and membrane fluidity of the cerebral cortex in aged male Wistar rats (65 and 91 weeks old) were affected by low-dose X-ray irradiation (100 cGy or under) compared with those in 7-week-old rats, to elucidate the mechanism of aging inhibition. The following results were obtained: Although radiation sensitivity was observed to decreases with age, low-dose irradiation changed the Mn-SOD activity, lipid peroxide level, and membrane protein fluidity parameter of the cerebral cortex in the age rats to be closer to those in the youth. These findings suggest that the increased SOD activity induced by low-dose irradiation enhances biomembrane functions, and that the decrease of lipid peroxide level enhances the membrane protein fluidity.

Probucol scavenged 1,1-diphenyl-2-picrylhydrazyl radicals and inhibited formation of thiobarbituric acid reactive substances

Probucol is suggested to have antioxidant properties. The direct scavenging action of probucol on hydroxyl radicals, superoxide and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were examined using electron spin resonance (ESR) spectrometry. Probucol scavenged DPPH radicals dose dependently but showed no effect on hydroxyl radicals and superoxide generated by Fenton reaction and by hypoxanthine-xanthine oxidase system, respectively. It inhibited the formation of thiobarbituric acid reactive substances (TBARS) in rat cortex homogenate induced by ascorbic acid and FeCl2 at low dose, but it increased TBARS formation at high doses. Probucol showed no effect on the carbon centered radicals. Iron injection into the rat cortex, which is an experimental model for traumatic epilepsy, increased TBARS level in the cortex, hippocampus, striatum and cerebellum, but pretreatment with probucol inhibited the increase in these brain parts except for the hippocampus. These results suggest that the antioxidant property of probucol is partly due to its free radical scavenging effect.

Metal ions affect neuronal membrane fluidity of rat cerebral cortex.

The effect of various metal ions on neuronal membrane fluidity was examined using 2-(14-carboxypropyl)-2-ethyl-4,4-dimethyl-3-oxazolidinyloxy, which has been used for the examination of membrane fluidity in hydrophobic areas by electron spin resonance spectrometry. Potassium, cobalt, calcium, magnesium, nickel, copper, ferric, and aluminium ions decreased the membrane fluidity while ferrous ions increased it at each high concentration. Sodium and zinc ions had no effect. Ethylenediaminetetraacetic acid decreased membrane fluidity at high concentrations. Nicardipine lowered membrane fluidity and flunarizine elevated it at each high concentration. There was no change in membrane fluidity by other calcium antagonists, nimodipine and nifedipine.

Regional excitatory and inhibitory amino acid levels in epileptic El mouse brain

Inbred mutant El mice are highly susceptible to convulsive seizures upon “tossing” stimulation. The levels of excitatory (e.g. glutamate and aspartate) and inhibitory amino acids [e.g. γ-aminobutyrate (GABA)] were examined in discrete regions of stimulated El mice [El(+)] non-stimulated El mice [El(-)] and ddY mice, which do not have convulsive disposition. In comparison with ddY, a general increased levels of aspartate, glutamate, glutamine, and taurine were detected in brain regions of El(-). The levels of GABA and glycine were almost the same in ddY and El(-). Compared to El(+), the levels of aspartate, glutamate, glutamine, and GABA in El(-) were either the same or higher. In the case of taurine and glycine, the levels in El(-) were either the same or lower than El(+). Alanine is special in that El(-) have a higher level than El(+) in hippocampus but lower in cerebellum. Furthermore, while marked changes were registered in several brain regions, none of the amino acids investigated showed any significant differences in the hypothalamus of three different groups of mice.

Free radical scavenging action of bio-catalyzer α·ϱ No. 11 (bio-normalyzer) and its by-product

Abstract Bio-catalyzer α·ϱ No. 11 (Bio-normalyzer) and its by-product are natural health products made by yeast fermentation of glucose, Carica papaya Linn., Pennisetum pupureum Schum., and Sechium edule Swartz. Their effects on free radicals were examined by electron spin resonance spectrometry using spin trapping agents 5,5-dimethyl-1-pyrroline-1-oxide (DMPO). It was observed that both Bio-catalyzer and its by-product scavenged 95% of DMPO-OH spin adducts (89 × 10 15 spins/ml) generated by FeSO 4 -H 2 O 2 -diethylene triamine pentaacetic acid system at 45.45 mg/ml each. Five percent of DMPO-O 2 − spin adducts (27 × 10 15 spins/ml) generated by hypoxanthine-xanthine oxidase system and 11% of 1,1-diphenyl-2-picrylhydrazyl radicals (7 × 10 15 spins/ml) were quenched using 25 mg/ml of Bio-catalyzer while 5% of superoxide and nil DPPH radicals were scavenged by its by-product. Vivo tests showed that oral administration of 1-g/kg body weight of Bio-catalyzer significantly inhibited thiobarbituric acid reactive substances formation, which is an index of lipid peroxidation, in the FeCl 3 -induced epileptic focus of rats. These findings suggest that Bio-catalyzer or its by-product may be useful health foods against neural lipid peroxidation, traumatic epilepsy and aging.