Curcuma zedoaria 50% methanol extracts increase adiponectin secretion by enhancing PPARγ mRNA expression in 3T3-L1 cells.

Curcuma zedoaria is a characteristic species of its genus that contains little to no curcuminoid. Here, we demonstrate that C. zedoaria extracts with 50% methanol increases adiponectin secretion into the media by enhancing PPARγ mRNA expression in 3T3-L1 cells. These results indicate that C. zedoaria may be useful for preventing/improving lifestyle-related diseases such as diabetes and atherosclerosis.

Classification of Japanese Pepper (Zanthoxylum piperitum DC.) from Different Growing Regions Based on Analysis of Volatile Compounds and Sensory Evaluation.

The Japanese pepper (Zanthoxylum piperitum DC.) is an attractive plant that is highly palatable and benefits human health. There are several lineages of pepper plants in Japan. However, the classification of each lineage by analyzing its volatile compounds and studies on the effects of differences in volatile compounds on human flavor perception have not been performed in detail. Herein, we conducted gas chromatography (GC) and GC/mass spectrometry (GC/MS) analysis of volatile compounds and sensory evaluation of flavor by an analytical panel using 10 commercially available dry powdered Japanese pepper products from different regions. GC and GC/MS analysis detected limonene, ß-phellandrene, citronellal, and geranyl acetate as the major volatile compounds of Japanese peppers. The composition of volatile compounds showed different characteristics depending on the growing regions, and cluster analysis of composition classified the products into five groups. The sensory evaluation classified the products into four groups, and the results of both classifications were in good agreement. Our results provide an important basis for proposing cooking and utilization methods that take advantage of the unique characteristics of each lineage based on scientific evidence.

Irbesartan attenuates atherosclerosis in Watanabe heritable hyperlipidemic rabbits: noninvasive imaging of inflammation by 18F-fluorodeoxyglucose positron emission tomography.

The purpose of this study was to assess the usefulness of 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET) in evaluating the antiatherogenic effects of irbesartan, an angiotensin II type 1 receptor blocker. Watanabe heritable hyperlipidemic rabbits were divided into the irbesartan-treated group (75 mg/kg/d; n  =  14) and the control group (n  =  14). After a 9-month treatment, rabbits underwent 18F-FDG PET. Using the aortic lesions, autoradiography and histologic examinations were performed. PET imaging clearly visualized the thoracic lesions of control rabbits and showed a significant decrease in the 18F-FDG uptake level of irbesartan-treated rabbits (78.8% of controls; p < .05). Irbesartan treatment significantly reduced the plaque size (43.1% of controls) and intraplaque macrophage infiltration level (48.1% of controls). The 18F-FDG uptake level in plaques positively correlated with the plaque size (r  =  .65, p < .05) and macrophage infiltration level (r  =  .57, p < .05). Noninvasive imaging by 18F-FDG PET is useful for evaluating the therapeutic effects of irbesartan and reflects inflammation, a key factor involved in the therapeutic effects.

Radiolabeled BMIPP for imaging hepatic fatty acid metabolism: evaluation of hepatic distribution and metabolism in mice at various metabolic statuses induced by fasting in comparison with palmitic acid.

Abnormalities in hepatic fatty acid metabolism are involved in various diseases. In order to clarify the use of 123I-15-(p-iodophenyl)-3(R,S)-methylpentadecanoic acid ([123I]BMIPP) for imaging hepatic fatty acid metabolism, we determined the hepatic distribution/metabolism of [125I]BMIPP in mice at various metabolic statuses induced by fasting, and compared the results with those of [1-(14)C]palmitic acid ([1-(14)C]PA). Fed or fasted (6, 12, and 24 hour-fasted) mice were intravenously injected with [125I]BMIPP or [1-(14)C]PA. Hepatic radioactivity was measured at 1 to 120 minutes after the injection (n  =  5 to 15/time points), and radioactive lipid metabolites were analyzed by thin-layer chromatography (n  =  3/time points). The areas under the curves (AUCs) were calculated. In mice given [125I]BMIPP, the hepatic radioactivity was increased with the fasting time (AUC: 35.1, 45.5, 57.6, and 59.0 [% injected dose (ID)/g/kg]▪min for fed, 6, 12, and 24 hour-fasted). Similar characteristic changes were observed in mice given [1-(14)C]PA (100.6, 101.0, 116.5, and 121.5 [%ID/g/kg]▪min). Metabolite analysis showed that the triglyceride-fraction was increased by fasting in both groups (5.7, 12.8, 32.0, and 37.9 [%ID/g/kg]▪min for [125I]BMIPP groups; 20.6, 39.2, 66.0, and 67.9 [%ID/g/kg]▪min for [1-(14)C]PA groups). Thus, [125I]BMIPP demonstrated the changes in hepatic fatty acid metabolism induced by fasting, indicating the potential of [123I]BMIPP for imaging hepatic fatty acid metabolism.

Screening and identification of disaccharides with insulin mimetic activity against L6 cells.

Insulin mimetics are considered as prospective anti-diabetic agents, and the disaccharide, neohesperidose, has been found to show insulin mimetic activity against L6 cells. We screened several other disaccharides for their insulin mimetic activity and identified three new insulin mimetic disaccharides.

Effects of feeding condition on the myocardial and hepatic accumulation of radioiodine-labeled BMIPP in mice.

OBJECTIVE: 123I-15-(p-iodophenyl)-3(R,S)-methylpentadecanoic acid ([123I]BMIPP), a fatty acid analog, is widely used for the diagnosis of cardiac diseases. Feeding condition is one of the important factors in the myocardial fatty acid uptake, which may also affect myocardial accumulation of [123I]BMIPP and image quality of [123I]BMIPP scintigraphy. However, the relationship between the myocardial accumulation of [123I]BMIPP and the feeding condition is not entirely clear. Therefore, we determined the myocardial accumulation of [125I]BMIPP in mice at various metabolic statuses induced by fasting in comparison with the hepatic accumulation. METHODS: Fed or fasted (6-, 12-, and 24-h fasted) mice were intravenously injected with [125I]BMIPP (35.2-75.0 kBq, 4 nmol). Radioactivities in the heart and liver were measured at 1, 5, 10, 30, 60, and 120 min after the injection (n = 5-15/time point for each group), and then, the heart-to-liver (H/L) ratios were calculated. RESULTS: The myocardial accumulation level of [125I]BMIPP in the fed group was almost the same as that in the 6-h-fasted group at each time point, although it was decreased by 12- and 24-h fasting. The H/L ratios of [125I]BMIPP accumulation level were significantly decreased by fasting (1.92 ± 0.22, 1.45 ± 0.13, 1.12 ± 0.13, and 0.91 ± 0.15 at 10 min, and 3.30 ± 0.62, 2.09 ± 0.35, 1.79 ± 0.34, and 1.27 ± 0.06 at 30 min after the injection, respectively, for the fed group and the 6-, 12-, and 24-h-fasted groups; p < 0.0001), largely owing to the increase in the hepatic accumulation level in the fasting groups. CONCLUSION: Although short-period (6 h) fasting did not affect the myocardial accumulation level of [125I]BMIPP, the hepatic accumulation level was increased. The present results indicate that the fed condition may provide higher-contrast images in myocardial [123I]BMIPP scintigraphy.

Elucidation of Japanese pepper (Zanthoxylum piperitum De Candolle) domestication using RAD-Seq.

Japanese pepper, Zanthoxylum piperitum, is native to Japan and has four well-known lineages (Asakura, Takahara, Budou, and Arima), which are named after their production area or morphology. Restriction-site associated DNA sequencing (RAD-Seq) was used to analyse 93 accessions from various areas, including these four lineages. Single nucleotide variant analysis was used to classify the plants into eight groups: the Asakura and Arima lineages each had two groups, the Takahara and Budou lineages each had one group, and two additional groups were present. In one Asakura group and two Arima groups, the plants were present in agricultural fields and mountains, thus representing the early stage of domestication of the Japanese pepper. The second Asakura lineage group was closely related to plants present in various areas, and this represents the second stage of domestication of this plant because, after early domestication, genetically related lineages with desirable traits spread to the periphery. These results demonstrate that domestication of Japanese pepper is ongoing. In addition, this study shows that spineless plants are polyphyletic, despite the spineless lineage being considered a subspecies of Japanese pepper.

In vitro uptake and metabolism of [14C]acetate in rabbit atherosclerotic arteries: biological basis for atherosclerosis imaging with [11C]acetate.

INTRODUCTION: Detection of vulnerable plaques is critically important for the selection of appropriate treatment and/or the prevention of atherosclerosis and ensuing cardiovascular diseases. In order to clarify the utility of [11C]acetate for atherosclerosis imaging, we determined the uptake and metabolism of acetate by in vitro studies using rabbit atherosclerotic arteries and [14C]acetate. METHODS: Rabbits were fed with a conventional (n=5) or a 0.5% cholesterol diet (n=6). One side of the iliac-femoral arteries was injured by a balloon catheter. Radioactivity levels in the iliac-femoral arteries were measured after incubation in DMEM containing [1-14C]acetate for 60 min (% dpm/mg tissue). Radioactive components in the homogenized arteries were partitioned into aqueous, organic, and residue fractions by the Folch method, and analyzed by thin-layer chromatography (TLC). RESULTS: The radioactivity level in the injured arteries of rabbits fed with the 0.5% cholesterol diet (atherosclerotic arteries) was significantly higher than that in either the non-injured or injured arteries of rabbits fed with the conventional diet (p<0.05) (% dpm/mg tissue: conventional diet groups; 0.022±0.005 and 0.024±0.007, cholesterol diet groups; 0.029±0.007 and 0.034±0.005 for non-injured and injured arteries). In metabolite analysis, most of the radioactivity was found in the aqueous fraction in each group (87.4-94.6% of total radioactivity in the arteries), and glutamate was a dominant component (67.4-69.7% of the aqueous fraction in the arteries). CONCLUSIONS: The level of [14C]acetate-derived radioactivity into the arteries was increased by balloon injury and the burden of a cholesterol diet. Water-soluble metabolites were the dominant components with radioactivity in the atherosclerotic lesions. These results provide a biological basis for imaging atherosclerotic lesions by PET using [11C]acetate.

Altered glucose metabolism and hypoxic response in alloxan-induced diabetic atherosclerosis in rabbits.

Diabetes mellitus accelerates atherosclerosis that causes most cardiovascular events. Several metabolic pathways are considered to contribute to the development of atherosclerosis, but comprehensive metabolic alterations to atherosclerotic arterial cells remain unknown. The present study investigated metabolic changes and their relationship to vascular histopathological changes in the atherosclerotic arteries of rabbits with alloxan-induced diabetes. Diabetic atherosclerosis was induced in rabbit ilio-femoral arteries by injecting alloxan (100 mg/kg), injuring the arteries using a balloon, and feeding with a 0.5% cholesterol diet. We histologically assessed the atherosclerotic lesion development, cellular content, pimonidazole positive-hypoxic area, the nuclear localization of hypoxia-inducible factor-1α, and apoptosis. We evaluated comprehensive arterial metabolism by performing metabolomic analyses using capillary electrophoresis-time of flight mass spectrometry. We evaluated glucose uptake and its relationship to vascular hypoxia using 18F-fluorodeoxyglucose and pimonidazole. Plaque burden, macrophage content, and hypoxic areas were more prevalent in arteries with diabetic, than non-diabetic atherosclerosis. Metabolomic analyses highlighted 12 metabolites that were significantly altered between diabetic and non-diabetic atherosclerosis. A half of them were associated with glycolysis metabolites, and their levels were decreased in diabetic atherosclerosis. The uptake of glucose evaluated as 18F-fluorodeoxyglucose in atherosclerotic lesions increased according to increased macrophage content or hypoxic areas in non-diabetic, but not diabetic rabbits. Despite profound hypoxic areas, the nuclear localization of hypoxia-inducible factor-1α decreased and the number of apoptotic cells increased in diabetic atherosclerotic lesions. Altered glycolysis metabolism and an impaired response to hypoxia in atherosclerotic lesions under conditions of insulin-dependent diabetes might be involved in the development of diabetic atherosclerosis.

Increased metabolite levels of glycolysis and pentose phosphate pathway in rabbit atherosclerotic arteries and hypoxic macrophage.

AIMS: Inflammation and possibly hypoxia largely affect glucose utilization in atherosclerotic arteries, which could alter many metabolic systems. However, metabolic changes in atherosclerotic plaques remain unknown. The present study aims to identify changes in metabolic systems relative to glucose uptake and hypoxia in rabbit atherosclerotic arteries and cultured macrophages. METHODS: Macrophage-rich or smooth muscle cell (SMC)-rich neointima was created by balloon injury in the iliac-femoral arteries of rabbits fed with a 0.5% cholesterol diet or a conventional diet. THP-1 macrophages stimulated with lipopolysaccharides (LPS) and interferon-γ (INFγ) were cultured under normoxic and hypoxic conditions. We evaluated comprehensive arterial and macrophage metabolism by performing metabolomic analyses using capillary electrophoresis-time of flight mass spectrometry. We evaluated glucose uptake and its relationship to vascular hypoxia using (18)F-fluorodeoxyglucose ((18)F-FDG) and pimonidazole, a marker of hypoxia. RESULTS: The levels of many metabolites increased in the iliac-femoral arteries with macrophage-rich neointima, compared with those that were not injured and those with SMC-rich neointima (glycolysis, 4 of 9; pentose phosphate pathway, 4 of 6; tricarboxylic acid cycle, 4 of 6; nucleotides, 10 of 20). The uptake of (18)F-FDG in arterial walls measured by autoradiography positively correlated with macrophage- and pimonidazole-immunopositive areas (r = 0.76, and r = 0.59 respectively; n = 69 for both; p<0.0001). Pimonidazole immunoreactivity was closely localized with the nuclear translocation of hypoxia inducible factor-1α and hexokinase II expression in macrophage-rich neointima. The levels of glycolytic (8 of 8) and pentose phosphate pathway (4 of 6) metabolites increased in LPS and INFγ stimulated macrophages under hypoxic but not normoxic condition. Plasminogen activator inhibitor-1 protein levels in the supernatant were closely associated with metabolic pathways in the macrophages. CONCLUSION: Infiltrative macrophages in atherosclerotic arteries might affect metabolic systems, and hypoxia but not classical activation might augment glycolytic and pentose phosphate pathways in macrophages.

Study of kaempferol glycoside as an insulin mimic reveals glycon to be the key active structure.

Diabetes mellitus is increasing in prevalence with patient numbers rising throughout the world. Current treatments for diabetes mellitus focus on control of blood glucose levels. Certain kinds of flavonoids or their glycosides stimulate cells to improve glucose uptake and lower blood glucose levels. We synthesized kaempferol 3-O-neohesperidoside (1), a naturally occurring substance present in Cyathea phalerata Mart., reported to mimic the action of insulin. Synthetic 1 promoted glucose uptake in the cultured cell line, L6. Further studies to determine the core structure responsible for this activity using synthetic compounds revealed neohesperidose to be the primary pharmacophore. These findings support the use of certain saccharides as a potential novel treatment for diabetes mellitus by replacing or supporting insulin.