Shizuka Tamaru

High-fat and high-cholesterol diet rapidly induces non-alcoholic steatohepatitis with advanced fibrosis in Sprague–Dawley rats

The development of fibrosis is considered an important phase in the progress of non‐alcoholic steatohepatitis (NASH) towards the end stage of liver disease, including cirrhosis. However, few small animal models can display NASH‐associated fibrosis. We aimed to establish a dietary model of NASH with rapid progression to fibrosis using genetically normal rats.

D-psicose, an epimer of D-fructose, favorably alters lipid metabolism in Sprague-Dawley rats.

D-Psicose, a C3 epimer of D-fructose, is known to lower body weight and adipose tissue weight and affect lipid metabolism. The precise mechanism remains unknown. It has been reported that D-psicose has a short half-life and is not metabolized in the body. To determine how D-psicose modifies lipid metabolism, rats were fed diets with or without 3% D-psicose for 4 weeks. Rats were decapitated without fasting every 6 h over a period of 24 h. Changes in serum and liver lipid levels, liver enzyme activity, and gene expression were quantified in experiment 1. Rats fed D-psicose had significantly lower serum insulin and leptin levels. Liver enzyme activities involved in lipogenesis were significantly lowered by the D-psicose diet, whereas gene expression of a transcriptional modulator of fatty acid oxidation was enhanced. In experiment 2, feeding the D-psicose diet gave significantly lower body weight (389 ± 3 vs 426 ± 6 g, p < 0.05) and food intake (23.8 ± 0.2 vs 25.7 ± 0.4 g/day, p < 0.05) compared to the control diet. Rats fed the D-psicose diet gave significantly higher energy expenditure in the light period and fat oxidation in the dark period compared to rats fed the control diet, whereas carbohydrate oxidation was lower. In summary, these results indicate that the D-psicose diet decreases lipogenesis, increases fatty acid oxidation, and enhances 24 h energy expenditure, leading to d-psicoses potential for weight management.

Hypoglycemic activity of Eriobotrya japonica seeds in type 2 diabetic rats and mice

The hypoglycemic effects of Eriobotrya japonica seeds were investigated in type 2 diabetic Otsuka Long-Evans Tokushima fatty (OLETF) rats and KK-Ay mice. The rats and mice were fed on a diet containing 10% powdered Eriobotrya japonica seeds with the coat intact for 4 months. Although the blood glucose concentration in the OLETF rats fed on the control diet without Eriobotrya japonica seeds was increased with time, the concentration in the OLETF rats fed on the diet with Eriobotrya japonica seeds was consistently low throughout the experimental period and was comparable to the level in Long-Evans Tokushima Otsuka (LETO) rats which are normal non-diabetic rats. Serum insulin was significantly lower in the OLETF rats fed on the Eriobotrya japonica seed diet than in those fed on the control diet at the termination of the experimental period. Eriobotrya japonica seeds suppressed the increment of blood glucose for 4 months and also effectively improved the glucose tolerance in the KK-Ay mice, these actions being mainly exerted by the ethanol extract of the seeds. These results suggest that Eriobotrya japonica seeds had a hypoglycemic property and the effect is attributable to the components extracted by ethanol.

Olive leaf extract prevents spontaneous occurrence of non-alcoholic steatohepatitis in SHR/NDmcr-cp rats

Aims: Oxidative stress may play an important role in the pathogenesis of non‐alcoholic steatohepatitis (NASH). Oleuropein, the active constituent of olive leaf, possesses anti‐oxidant, hypoglycaemic, and hypolipidaemic activities. We aimed to investigate the preventive effects of olive leaf extract on hepatic fat accumulation in a rat model of NASH. Methods: Spontaneously hypertensive/NIH‐corpulent rats were fed a diet of AIN‐93G with or without olive leaf extract (500, 1000, 2000 mg/kg diet, and control; 5 rats each) for 23 weeks. Serological and histopathological findings, anti‐oxidative activity, and the alteration of fatty acid synthesis in the liver were evaluated. Results: Histopathologically, a diet of AIN‐93G containing more than 1000 mg/kg olive leaf extract had a preventive effect for the occurrence of NASH. Thioredoxin‐1 expression in the liver was more evident in rats fed this diet, and 4‐hydroxynonenal expression in the liver was less evident in these rats. There were no significant differences in the activities of hepatic carnitine palmitoyltransferase, fatty acid synthase, malic enzyme, and phosphatidic acid phosphohydrolase among the groups. Conclusions: Our data suggest that olive leaf extract may help prevent NASH, presumably through its anti‐oxidative activity.

Theflavins and theasinensin A derived from fermented tea have antihyperglycemic and hypotriacylglycerolemic effects in KK-Ay mice and Sprague-Dawley rats

Although tea polyphenols are reported to improve serum glucose and lipid levels by inhibiting amylase activity and reducing lipid absorption, in vivo data are lacking. We evaluated in vivo the antihyperglycemic and hypotriacylglycerolemic effects of theaflavins (TFs) and theasinensin A (TSA) refined from fermented tea to purities of 12 and 59%, respectively. Feeding male KK-A(y) mice diets with 0.1% TFs or TSA for 6 weeks reduced serum glucose levels by >30% compared to a control diet. Rats fed diets containing 0.2% TFs or TSA for 4 weeks had higher fecal fat excretion and 33% lower hepatic triacylglycerol; hepatic fatty acid synthase activity was not affected. Oral administration of TFs or TSA reduced the increase in serum triacylglycerol after an oral bolus of a fat emulsion. These results indicate TFs and TSA induce antihyperglycemic responses in diabetic mice and are hypotriacylglycerolemic in rats by suppressing intestinal fat absorption.

Hypotriacylglycerolemic and Antiobesity Properties of a New Fermented Tea Product Obtained by Tea-Rolling Processing of Third-Crop Green Tea (Camellia sinensis) Leaves and Loquat (Eriobotrya japonica) Leaves

We manufactured a new fermented tea by tea-rolling processing of third-crop green tea (Camellia sinensis) leaves and loquat (Eriobotrya japonica) leaves. The mixed fermented tea extract inhibited pancreatic lipase activity in vitro, and effectively suppressed postprandial hypertriacylglycerolemia in rats. Rats fed a diet containing 1% freeze-dried fermented tea extract for 4 weeks had a significantly lower liver triacylglycerol concentration and white adipose tissue weight than those fed the control diet lacking fermented tea extract. The activity of fatty acid synthase in hepatic cytosol markedly decreased in the fermented tea extract group as compared to the control group. The serum and liver triacylglycerol- and body fat-lowering effects of the mixed fermented tea extract were strong relative to the level of dietary supplementation. These results suggest that the new fermented tea product exhibited hypotriacylglycerolemic and antiobesity properties through suppression of both liver fatty acid synthesis and postprandial hypertriacylglycerolemia by inhibition of pancreatic lipase.

Transepithelial transport of theasinensins through Caco-2 cell monolayers and their absorption in Sprague-Dawley rats after oral administration.

The aim of this study is to illustrate the in vivo and in vitro absorption of theasinensins B and A that are (-)-epigallocatechin-3-O-gallate (EGCG)-(-)-epigallocatechin (EGC) dimer and EGCG dimer, respectively, and their transport pathway across the intestinal membrane. Our animal study by a single oral administration to rats demonstrated the intact absorption of theasinensins into the blood system, which was estimated to be a >10-fold lower absorption amount than EGCG. The in vitro absorption study indicated that theasinensins can be transported across Caco-2 cell monolayers, while their permeability coefficients were also >10-fold lower than those of EGCG and EGC. Transport experiments using cytochalasin D or quercetin as a tight junction (TJ) modulator and a non-saturable permeation revealed that theasinensins were transported across Caco-2 cells in a TJ paracellular diffusion route. In conclusion, the dimers of condensed catechins, theasinensins B and A, can be absorbed intact into rat blood and transported across Caco-2 cell monolayers probably through a TJ paracellular pathway.

Polyphenol composition of a functional fermented tea obtained by tea-rolling processing of green tea and loquat leaves

Phenolic constituents of a new functional fermented tea produced by tea-rolling processing of a mixture (9:1) of tea leaves and loquat leaves were examined in detail. The similarity of the phenolic composition to that of black tea was indicated by high-performance liquid chromatography comparison with other tea products. Twenty-five compounds, including three new catechin oxidation products, were isolated, and the structures of the new compounds were determined to be (2R)-2-hydroxy-3-(2,4,6-trihydroxyphenyl)-1-(3,4,5-trihydroxyphenyl)-1-propanone 2-O-gallate, dehydrotheasinensin H, and acetonyl theacitrin A by spectroscopic methods. In addition, theacitrinin A and theasinensin H were obtained for the first time from commercial tea products. Isolation of these new and known compounds confirms that reactions previously demonstrated by in vitro model experiments actually occur when fresh tea leaves are mechanically distorted and bruised during the production process.

Hypotriglyceridemic potential of fermented mixed tea made with third-crop green tea leaves and camellia (Camellia japonica) leaves in Sprague-Dawley rats.

Fermented mixed tea made with third-crop green tea leaves and camellia leaves by a tea-rolling process has been developed. The objective of this study was to investigate hypotriglyceridemic potential of the mixed tea in rats. The mixed tea contained theasinensins and theaflavins. Rats fed the mixed tea extract at the level of 1% exerted significantly lower body weight and adipose tissue weight compared to animals fed third-crop green tea or camellia tea extract alone for 4 weeks. Serum and hepatic triglyceride was significantly and dose-dependently decreased by the mixed tea. This decrease was associated with lowered lipogenic enzyme activities in the liver. Furthermore, an oral administration of 4 or 8% of the mixed tea extract followed by fat emulsion suppressed the increment of serum triglyceride level. These results suggest that the mixed tea has hypotriglyceridemic action, partially via delaying triglyceride absorption in the small intestine and repressing hepatic lipogenic enzymes.

Insoluble Fiber in Young Barley Leaf Suppresses the Increment of Postprandial Blood Glucose Level by Increasing the Digesta Viscosity

Barley (Hordeum vulgare L.) is a well-known cereal plant. Young barley leaf is consumed as a popular green-colored drink, which is named “Aojiru” in Japan. We examined the effects of barley leaf powder (BLP) and insoluble fibers derived from BLP on postprandial blood glucose in rats and healthy Japanese volunteers. BLP and insoluble fibers derived from BLP suppressed the increment of postprandial blood glucose levels in rats (P < 0.01), and increased the viscosity of their digesta. The insoluble fibers present in BLP might play a role in controlling blood glucose level by increasing digesta viscosity. In human, BLP suppressed the increment of postprandial blood glucose level only in those which exhibited higher blood glucose levels after meals (P < 0.01). BLP might suppress the increment of postprandial blood glucose level by increasing digesta viscosity in both of rats and humans who require blood glucose monitoring.