Tomonori Unno

Inhibiting Effects of Theanine on Caffeine Stimulation Evaluated by EEG in the Rat

In this study, the inhibiting action of theanine on the excitation by caffeine at the concentration regularly associated with drinking tea was investigated using electroencephalography (EEG) in rats. First, the stimulatory action by caffeine i.v. administration at a level higher than 5 μmol/kg (0.970 mg/kg) b.w. was shown by means of brain wave analysis, and this level was suggested as the minimum dose of caffeine as a stimulant. Next, the stimulatory effects of caffeine were inhibited by an i.v. administration of theanine at a level higher than 5 μmol/kg (0.781 mg/kg) b.w., and the results suggested that theanine has an antagonistic effect on caffeine’s stimulatory action at an almost equivalent molar concentration. On the other hand, the excitatory effects were shown in the rat i.v. administered 1 and 2 μmol/kg (0.174 and 0.348 mg/kg) b.w. of theanine alone. These results suggested two effects of theanine, depending on its concentration.

Protective effect of γ-glutamylethylamide (theanine) on ischemic delayed neuronal death in gerbils

We examined the protective effect of γ-glutamylethylamide (theanine) on ischemic delayed neuronal death in field CA1 of the gerbil hippocampus. One microliter of theanine from each three concentrations (50, 125 and 500 μM) was administered through the lateral ventricle 30 min before ischemia. Transient forebrain ischemia was induced by bilateral occlusion of the common carotid arteries for 3 min under careful control of brain temperature at approximately 37°C. Seven days after ischemia, the number of intact CA1 neurons in the hippocampus was assessed. Ischemia-induced neuronal death in hippocampal CA1 region was significantly prevented in a dose-dependent manner in the theanine-pretreated groups. These findings indicate that theanine might be useful clinically for preventing ischemic neuronal damage.

Effect of tea catechins on postprandial plasma lipid responses in human subjects

Epidemiological surveys suggest that a higher intake of tea may be associated with a lower risk of CHD. There is accumulating evidence that postprandial lipaemia makes a substantial contribution to the incidence of CHD. Our aim was, therefore, to evaluate the effect of tea catechins (major ingredients in green tea) on postprandial lipid responses in human subjects after the consumption of test meals. In a randomized triple-crossover design, nine male subjects with mild or borderline hypertriacylglycerolaemia consumed 10 (control), 224 (moderate dose) and 674 mg (high dose) of the assigned tea catechins three times each along with a standardized light meal consisting of a piece of bread spread with 20 g butter. Plasma lipids were measured in the fasting state and 1, 2, 3, 4 and 6 h after consuming the light meal. Results showed that, compared with the control, moderate and high doses of tea catechins reduced the incremental area under the plasma triacylglycerol curves by 15.1 and 28.7%, respectively. Next, the rapid elevation of remnant-like particle cholesterol was significantly inhibited by a high dose of tea catechins 2 h after consuming the light meal (P<0.01). In the range of tea catechin dosages, no significant differences were observed in the postprandial responses for plasma total cholesterol or NEFA at any time point. In conclusion, this trial demonstrated that tea catechins attenuated the postprandial increase in plasma triacylglycerol levels following a fat load. These results may provide evidence for one of the possible mechanisms involved in lowering the incidence of CVD, and may prove useful in further studies on the beneficial health effects of tea drinking.

Scavenging effect of tea catechins and their epimers on superoxide anion radicals generated by a hypoxanthine and xanthine oxidase system

Catechins are a major group of polyphenolic compounds contained in abundance in green tea. Using electron spin resonance spectroscopy along with a spin-trapping agent, the scavenging effect of tea catechins and their corresponding epimers against superoxide anion radicals generated by a hypoxanthine and xanthine oxidase reaction system was evaluated. The presence of 3′,4′,5′-trihydroxyl groups attached to the B-ring of the flavan skeleton of tea catechins elevated their radical-scavenging efficiency in comparison to those with 3′,4′-dihydroxyl groups. There were no significant differences between the four dominant tea catechins and their corresponding epimers with regard to radical-scavenging ability. Under the different spin-trapping agent concentrations, the sigmoid curves of reducing spin-trapping adducts produced by tea catechins were shifted leftward, suggesting that a likely possible action of tea catechins is to scavenge superoxide anion radicals directly, not to inhibit the function of xanthine oxidase. Although caffeine is also known as a major ingredient of tea, its superoxide anion radical-scavenging effect was much weaker than that of the catechin family. It is concluded that tea catechins and their epimers serve as powerful antioxidants for directly eliminating superoxide anion radicals, and may be useful in the prevention of diseases relating to in vivo oxidative stress. © 2000 Society of Chemical Industry

Antioxidant activity of various teas against free radicals and LDL oxidation

Tea is a widely consumed beverage throughout the world. We assessed the antioxidant activity of six teas, including the aqueous extracts of green tea and oolong tea (Camellia sinensis), tochu (Eucommia ulmoides), Gymnema sylvestre, Japanese mugwort (Artemisia princeps), and barley (Hordeum vulgare), against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and LDL oxidation, and examined the association of LDL oxidizability with the plasma catechin levels in 10 healthy volunteers with a single dose of 5 g green tea powder. In vitro, the inhibitory effects of DPPH radicals and LDL oxidation were found to be strongest in the extract of green tea and weakest in that of barley. After the ingestion of green tea powder, the lag time increased from basal 52.2±4.1 to 60.3±4.2 min at 1 h and 59.5±4.1 min at 2 h, and then returned to the baseline lag time (51.9±1.4 at 4 h and 52.1±4.7 min at 6 h). Regarding the plasma catechin levels, epigallocatechingallate and epicatechingallate significantly increased from basal 3.7±1.3 and 0.8±0.8 ng/mL to 65.7±11.6 and 54.6±12.6 ng/mL at 1 h, and 74.4±18.6 and 49.4±7.1 ng/mL at 2 h, respectively. Green tea therefore showed the strongest antioxidant activity among the six different teas, and the inhibitory effects of green tea on LDL oxidation depended on the plasma catechin levels.

Heat-Epimerized Tea Catechins Have the Same Cholesterol-Lowering Activity as Green Tea Catechins in Cholesterol-Fed Rats

Tea catechins are known to be epimerized by heat treatment. The effect of heat-epimerized tea catechins on serum cholesterol concentration was compared with that of green tea catechins. Our observations strongly suggest that both tea catechins and heat-epimerized tea catechins lower serum cholesterol concentration by inhibiting cholesterol absorption in the intestine. There was no differential effect between the two catechin preparations.

Electron spin resonance spectroscopic evaluation of scavenging activity of tea catechins on superoxide radicals generated by a phenazine methosulfate and NADH system

Abstract The scavenging effects of tea catechins on superoxide radicals (·O2−) generated non-enzymatically by a phenazine methosulfate (PMS) and reduced β-nicotinamide adenine dinucleotide (NADH) system, was studied using an electron spin resonance (ESR) spectrometer along with a spin-trapping agent, 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO). The presence of 3′, 4′, 5′-trihydroxyl groups attached to the B-ring of the flavan skeleton enhanced the radical scavenging efficiency displayed by the catechin family in comparison to those with 3′, 4′-dihydroxyl groups, and the insertion of a galloyl moiety into three positions of the C-ring exerted a synergistic impact on ·O2− scavenging activity. Catechin constituents accounted for 86% of the total ·O2− scavenging capacity of green tea extract, with a particularly high contribution ascribed to (−)-epigallocatechin gallate [(−)-EGCG] (48%) and (−)-epigallocatechin [(−)-EGC] (26%).

Theanine, γ-glutamylethylamide, is metabolized by renal phosphate-independent glutaminase

Abstract The distribution of theanine-degrading activity in Wistar rats was examined and this activity was detected only in the kidney. Judging from polyacrylamide gel electrophoresis, theanine-degrading enzyme from rat kidney was purified almost to homogeneity. Theanine-degrading activity was co-purified with glutaminase activity, and the relative activity for theanine was about 85% of that for l -glutamine throughout purification. Substrate specificity of purified enzyme preparation coincided well with the data of phosphate-independent glutaminase [EC 3.5.1.2], which had been previously reported. It was very curious that γ-glutamyl methyl and ethyl esters were more effectively hydrolyzed than theanine and l -glutamine, in view of relative activity and K m value. It was suggested that γ-glutamyl moiety in theanine molecule was transferred to form γ-glutamylglycylglycine with relative ease in the presence of glycylglycine. On the other hand, purified phosphate-dependent glutaminase did not show theanine-degrading activity at all. Thus, it was concluded that theanine was hydrolyzed by phosphate-independent glutaminase in kidney and suggested that, as for the metabolic fate of theanine, its glutamyl moiety might be transferred by means of γ-glutamyl transpeptidase reaction to other peptides in vivo.

Antioxidative Activity of Water Extracts of Lagerstroemia speciosa Leaves

In order to develop naturally occurring antioxidants from edible plants, the antioxidative effect of hot water extracts of Lagerstroemia speciosa leaves, known by the Tagalog name of banaba in the Phillipines, was studied. The content of tannin in banaba extract was 36.8% in dry weight. Banaba extract showed strong antioxidative activity in a linoleic acid autoxidation system. Banaba extract was found to have a potent radical scavenging action on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and superoxide radicals (O(2-) ) generated by a hypoxanthine (HPX)/xanthine oxidase (XOD) system. In vitro lipid peroxidation of rat liver homogenate induced by tert-butyl hydroperoxide (BHP) was inhibited by the addition of banaba extract in a dose-dependent manner. From these results, banaba extract was demonstrated to be useful as an antioxidant or free radical scavenger to protect biological systems against oxidative stress.

Dose-Dependent Suppression of Tea Catechins with a Galloyl Moiety on Postprandial Hypertriglyceridemia in Rats

Tea has long been believed to be a healthy beverage, and its beneficial effects are almost all attributed to catechins. The effect of catechins on postprandial hypertriglyceridemia in rats was investigated in this study. A lipid emulsion administered orally to rats with (−)-epigallocatechin gallate at a dose of 100 mg/kg resulted in the increase in plasma triacylglycerol being significantly inhibited after 1 and 2 h compared to the case without (−)-epigallocatechin gallate. The effect of (−)-epigallocatechin was weaker than that of (−)-epigallocatechin gallate. A tea extract (THEA-FLAN 90S), mainly composed of catechins with a galloyl moiety, dose-dependently suppressed postprandial triacylglycerol after the administration of a lipid emulsion at doses of 50–200 mg/kg. The administration of the tea extract alone at a dose of 200 mg/kg had no effect on the plasma triacylglycerol level. These results strongly suggest that catechins with a galloyl moiety would be promising agents for suppressing dietary fat absorption through the small intestine.