26 results on '"Miyamoto, L."'
Search Results
2. Changes in intestinal microbiota and biochemical parameters in patients with inflammatory bowel disease and irritable bowel syndrome induced by the prolonged addition of soluble fibers to usual drug therapy.
- Author
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Watanabe H, Inoue T, Miyamoto L, Ono Y, Matsumoto K, Takeda M, and Tsuchiya K
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- Humans, Male, Female, Adult, Middle Aged, Inflammatory Bowel Diseases drug therapy, Feces microbiology, Feces chemistry, Fatty Acids, Volatile analysis, Fatty Acids, Volatile metabolism, Gastrointestinal Microbiome drug effects, Irritable Bowel Syndrome drug therapy, Irritable Bowel Syndrome microbiology, Dietary Fiber administration & dosage, Mannans administration & dosage, Plant Gums administration & dosage, Galactans administration & dosage
- Abstract
Objectives: Partially hydrolyzed guar gum (PHGG) is a soluble dietary fiber;in addition to improving bowel movements, it maintains intestinal health by producing short-chain fatty acids. However, majority of clinical studies on PHGG have been concluded within a month and excluded usual drug therapy. Hence, this study aimed to determine the effects of long-term consumption of PHGG, in combination with drug therapy, on gut bacteria ratios, laboratory values for inflammatory response, and fecal characteristics., Methods and Results: The study was performed in patients with irritable bowel syndrome (IBS), Crohn's disease (CD), and ulcerative colitis (UC), by the administration of PHGG for six months while they continued their usual treatment. PHGG treatment caused significant changes in patients with IBS, including an increase in the abundance of short-chain fatty acid-producing bacteria, a significant decrease in Bacteroides abundance, and normalization of the Bristol scale of stool. In patients with UC, non-significant normalization of soft stools and decrease in fecal calprotectin were observed. Adverse events were not observed in any of the groups., Conclusion: Thus, it would be beneficial to include PHGG in the usual drug therapies of patients with IBS. J. Med. Invest. 71 : 121-128, February, 2024.
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- 2024
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3. Indigo Leaves-Induced Pulmonary Arterial Remodeling without Right Ventricular Hypertrophy in Rats.
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Tsunematsu H, Imanishi M, Uemura Y, Higaki Y, Morisaki M, Katsura A, Miyamoto L, Funamoto M, Ichimura-Shimizu M, Horinouchi Y, Ikeda Y, Tsuneyama K, and Tsuchiya K
- Subjects
- Animals, Male, Hypertension, Pulmonary chemically induced, Hypertension, Pulmonary metabolism, Rats, Endothelial Cells drug effects, Endothelial Cells metabolism, Lung drug effects, Lung pathology, Lung metabolism, Plant Leaves, Pulmonary Artery drug effects, Pulmonary Artery metabolism, Pulmonary Artery pathology, Endothelin-1 metabolism, Vascular Remodeling drug effects, Hypertrophy, Right Ventricular metabolism, Hypertrophy, Right Ventricular physiopathology, Rats, Sprague-Dawley
- Abstract
Indigo naturalis (IN), derived from the leaves of the indigo plant, is a traditional Chinese medicine that has historically been used for its anti-inflammatory properties in the treatment of various diseases, including ulcerative colitis (UC). However, long-term use of IN in UC patients is incontrovertibly associated with the onset of pulmonary arterial hypertension (PAH). To investigate the mechanisms by which IN induces PAH, we focused on the raw material of IN, indigo leaves (IL). Only the condition of long-term chronic (6 months) and high-dose (containing 5% IL in the control diet) administration of IL induced medial thickening in the pulmonary arteries without right ventricular hypertrophy in our rat model. IL administration for a month did not induce pulmonary arterial remodeling but increased endothelin-1 (ET-1) expression levels within endothelial cell (EC) layers in the lungs. Gene Expression Omnibus analysis showed that ET-1 is a key regulator of PAH and that the IL component indican and its metabolite IS induced ET-1 mRNA expression via reactive oxygen species-dependent mechanism. We identified the roles of indican and IS in ET-1 expression in ECs, which were linked to pulmonary arterial remodeling in an animal model.
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- 2024
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4. CA9 and PRELID2; hypoxia-responsive potential therapeutic targets for pancreatic ductal adenocarcinoma as per bioinformatics analyses.
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Imanishi M, Inoue T, Fukushima K, Yamashita R, Nakayama R, Nojima M, Kondo K, Gomi Y, Tsunematsu H, Goto K, Miyamoto L, Funamoto M, Denda M, Ishizawa K, Otaka A, Fujino H, Ikeda Y, and Tsuchiya K
- Subjects
- Humans, Carbonic Anhydrase IX genetics, Carbonic Anhydrase IX metabolism, Antigens, Neoplasm genetics, Antigens, Neoplasm metabolism, Antigens, Neoplasm therapeutic use, Hypoxia metabolism, RNA, Small Interfering, Computational Biology, Pancreatic Neoplasms, Carcinoma, Pancreatic Ductal drug therapy, Carcinoma, Pancreatic Ductal genetics, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms genetics
- Abstract
A strong hypoxic environment has been observed in pancreatic ductal adenocarcinoma (PDAC) cells, which contributes to drug resistance, tumor progression, and metastasis. Therefore, we performed bioinformatics analyses to investigate potential targets for the treatment of PDAC. To identify potential genes as effective PDAC treatment targets, we selected all genes whose expression level was related to worse overall survival (OS) in The Cancer Genome Atlas (TCGA) database and selected only the genes that matched with the genes upregulated due to hypoxia in pancreatic cancer cells in the dataset obtained from the Gene Expression Omnibus (GEO) database. Although the extracted 107 hypoxia-responsive genes included the genes that were slightly enriched in angiogenic factors, TCGA data analysis revealed that the expression level of endothelial cell (EC) markers did not affect OS. Finally, we selected CA9 and PRELID2 as potential targets for PDAC treatment and elucidated that a CA9 inhibitor, U-104, suppressed pancreatic cancer cell growth more effectively than 5-fluorouracil (5-FU) and PRELID2 siRNA treatment suppressed the cell growth stronger than CA9 siRNA treatment. Thus, we elucidated that specific inhibition of PRELID2 as well as CA9, extracted via exhaustive bioinformatic analyses of clinical datasets, could be a more effective strategy for PDAC treatment., (Copyright © 2023 The Authors. Production and hosting by Elsevier B.V. All rights reserved.)
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- 2023
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5. Diphenhydramine may be a preventive medicine against cisplatin-induced kidney toxicity.
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Hamano H, Ikeda Y, Goda M, Fukushima K, Kishi S, Chuma M, Yamashita M, Niimura T, Takechi K, Imanishi M, Zamami Y, Horinouchi Y, Izawa-Ishizawa Y, Miyamoto L, Ishizawa K, Fujino H, Tamaki T, Aihara KI, and Tsuchiya K
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- Animals, Apoptosis, Cisplatin toxicity, Diphenhydramine metabolism, Diphenhydramine pharmacology, Diphenhydramine therapeutic use, Humans, Kidney metabolism, Mice, Oxidative Stress, Retrospective Studies, Acute Kidney Injury chemically induced, Acute Kidney Injury drug therapy, Acute Kidney Injury prevention & control, Antineoplastic Agents toxicity
- Abstract
Cisplatin is widely used as an anti-tumor drug for the treatment of solid tumors. Unfortunately, it causes kidney toxicity as a critical side effect, limiting its use, given that no preventive drug against cisplatin-induced kidney toxicity is currently available. Here, based on a repositioning analysis of the Food and Drug Administration Adverse Events Reporting System, we found that a previously developed drug, diphenhydramine, may provide a novel treatment for cisplatin-induced kidney toxicity. To confirm this, the actual efficacy of diphenhydramine was evaluated in in vitro and in vivo experiments. Diphenhydramine inhibited cisplatin-induced cell death in kidney proximal tubular cells. Mice administered cisplatin developed kidney injury with significant dysfunction (mean plasma creatinine: 0.43 vs 0.15 mg/dl) and showed augmented oxidative stress, increased apoptosis, elevated inflammatory cytokines, and MAPKs activation. However, most of these symptoms were suppressed by treatment with diphenhydramine. Furthermore, the concentration of cisplatin in the kidney was significantly attenuated in diphenhydramine-treated mice (mean platinum content: 70.0 vs 53.4 μg/g dry kidney weight). Importantly, diphenhydramine did not influence or interfere with the anti-tumor effect of cisplatin in any of the in vitro or in vivo experiments. In a selected cohort of 98 1:1 matched patients from a retrospective database of 1467 patients showed that patients with malignant cancer who had used diphenhydramine before cisplatin treatment exhibited significantly less acute kidney injury compared to ones who did not (6.1 % vs 22.4 %, respectively). Thus, diphenhydramine demonstrated efficacy as a novel preventive medicine against cisplatin-induced kidney toxicity., (Copyright © 2020 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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6. Deletion of H-ferritin in macrophages alleviates obesity and diabetes induced by high-fat diet in mice.
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Ikeda Y, Watanabe H, Shiuchi T, Hamano H, Horinouchi Y, Imanishi M, Goda M, Zamami Y, Takechi K, Izawa-Ishizawa Y, Miyamoto L, Ishizawa K, Aihara KI, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Apoferritins genetics, Diabetes Mellitus etiology, Male, Mice, Mice, Knockout, Obesity etiology, Random Allocation, Apoferritins metabolism, Diabetes Mellitus metabolism, Diabetes Mellitus therapy, Diet, High-Fat adverse effects, Macrophages metabolism, Obesity metabolism, Obesity therapy
- Abstract
Aims/hypothesis: Iron accumulation affects obesity and diabetes, both of which are ameliorated by iron reduction. Ferritin, an iron-storage protein, plays a crucial role in iron metabolism. H-ferritin exerts its cytoprotective action by reducing toxicity via its ferroxidase activity. We investigated the role of macrophage H-ferritin in obesity and diabetes., Methods: Conditional macrophage-specific H-ferritin (Fth, also known as Fth1) knockout (LysM-Cre Fth KO) mice were used and divided into four groups: wild-type (WT) and LysM-Cre Fth KO mice with normal diet (ND), and WT and LysM-Cre Fth KO mice with high-fat diet (HFD). These mice were analysed for characteristics of obesity and diabetes, tissue iron content, inflammation, oxidative stress, insulin sensitivity and metabolic measurements. RAW264.7 macrophage cells were used for in vitro experiments., Results: Iron concentration reduced, and mRNA expression of ferroportin increased, in macrophages from LysM-Cre Fth KO mice. HFD-induced obesity was lower in LysM-Cre Fth KO mice than in WT mice at 12 weeks (body weight: KO 34.6 ± 5.6 g vs WT 40.1 ± 5.2 g). mRNA expression of inflammatory cytokines and infiltrated macrophages and oxidative stress increased in the adipose tissue of HFD-fed WT mice, but was not elevated in HFD-fed LysM-Cre Fth KO mice. However, WT mice fed an HFD had elevated iron concentration in adipose tissue and spleen, which was not observed in LysM-Cre Fth KO mice fed an HFD (adipose tissue [μmol Fe/g protein]: KO 1496 ± 479 vs WT 2316 ± 866; spleen [μmol Fe/g protein]: KO 218 ± 54 vs WT 334 ± 83). Moreover, HFD administration impaired both glucose tolerance and insulin sensitivity in WT mice, which was ameliorated in LysM-Cre Fth KO mice. In addition, energy expenditure, mRNA expression of thermogenic genes, and body temperature were higher in KO mice with HFD than WT mice with HFD. In vitro experiments showed that iron content was reduced, and lipopolysaccharide-induced Tnf-α (also known as Tnf) mRNA upregulation was inhibited in a macrophage cell line transfected with Fth siRNA., Conclusions/interpretation: Deletion of macrophage H-ferritin suppresses the inflammatory response by reducing intracellular iron levels, resulting in the prevention of HFD-induced obesity and diabetes. The findings from this study highlight macrophage iron levels as a potential therapeutic target for obesity and diabetes.
- Published
- 2020
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7. Renoprotective effects of a factor Xa inhibitor: fusion of basic research and a database analysis.
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Horinouchi Y, Ikeda Y, Fukushima K, Imanishi M, Hamano H, Izawa-Ishizawa Y, Zamami Y, Takechi K, Miyamoto L, Fujino H, Ishizawa K, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Cells, Cultured, Humans, Inflammation pathology, Inflammation prevention & control, Kidney Diseases pathology, Macrophages metabolism, Macrophages pathology, Male, Mice, Mice, Inbred C57BL, Nephritis, Interstitial pathology, Ureteral Obstruction pathology, Databases, Factual, Factor Xa Inhibitors pharmacology, Kidney Diseases prevention & control, Macrophages drug effects, Nephritis, Interstitial drug therapy, Pyridines pharmacology, Thiazoles pharmacology, Ureteral Obstruction drug therapy
- Abstract
Renal tubulointerstitial injury, an inflammation-associated condition, is a major cause of chronic kidney disease (CKD). Levels of activated factor X (FXa), a blood coagulation factor, are increased in various inflammatory diseases. Therefore, we investigated the protective effects of an FXa inhibitor against renal tubulointerstitial injury using unilateral ureteral obstruction (UUO) mice (a renal tubulointerstitial fibrosis model) and the Food and Drug Administration Adverse Events Reporting System (FAERS) database. The renal expression levels of FX and the FXa receptors protease-activated receptor (PAR)-1 and PAR-2 were significantly higher in UUO mice than in sham-operated mice. UUO-induced tubulointerstitial fibrosis and extracellular matrix expression were suppressed in UUO mice treated with the FXa inhibitor edoxaban. Additionally, edoxaban attenuated UUO-induced macrophage infiltration and inflammatory molecule upregulation. In an analysis of the FAERS database, there were significantly fewer reports of tubulointerstitial nephritis for patients treated with FXa inhibitors than for patients not treated with inhibitors. These results suggest that FXa inhibitors exert protective effects against CKD by inhibiting tubulointerstitial fibrosis.
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- 2018
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8. The uremic toxin indoxyl sulfate interferes with iron metabolism by regulating hepcidin in chronic kidney disease.
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Hamano H, Ikeda Y, Watanabe H, Horinouchi Y, Izawa-Ishizawa Y, Imanishi M, Zamami Y, Takechi K, Miyamoto L, Ishizawa K, Tsuchiya K, and Tamaki T
- Subjects
- Adenine toxicity, Animals, Disease Models, Animal, Hep G2 Cells, Hepcidins genetics, Humans, Male, Mice, Mice, Inbred C57BL, Oxidative Stress drug effects, Renal Insufficiency, Chronic chemically induced, Renal Insufficiency, Chronic pathology, Gene Expression Regulation drug effects, Hepcidins metabolism, Indican pharmacology, Iron metabolism, Renal Insufficiency, Chronic metabolism
- Abstract
Background: Hepcidin secreted by hepatocytes is a key regulator of iron metabolism throughout the body. Hepcidin concentrations are increased in chronic kidney disease (CKD), contributing to abnormalities in iron metabolism. Levels of indoxyl sulfate (IS), a uremic toxin, are also elevated in CKD. However, the effect of IS accumulation on iron metabolism remains unclear., Methods: We used HepG2 cells to determine the mechanism by which IS regulates hepcidin concentrations. We also used a mouse model of adenine-induced CKD. The CKD mice were divided into two groups: one was treated using AST-120 and the other received no treatment. We examined control mice, CKD mice, CKD mice treated using AST-120 and mice treated with IS via drinking water., Results: In the in vitro experiments using HepG2 cells, IS increased hepcidin expression in a dose-dependent manner. Silencing of the aryl hydrocarbon receptor (AhR) inhibited IS-induced hepcidin expression. Furthermore, IS induced oxidative stress and antioxidant drugs diminished IS-induced hepcidin expression. Adenine-induced CKD mice demonstrated an increase in hepcidin concentrations; this increase was reduced by AST-120, an oral adsorbent of the uremic toxin. CKD mice showed renal anemia, decreased plasma iron concentration, increased plasma ferritin and increased iron content in the spleen. Ferroportin was decreased in the duodenum and increased in the spleen. These changes were ameliorated by AST-120 treatment. Mice treated by direct IS administration showed hepatic hepcidin upregulation., Conclusions: IS affects iron metabolism in CKD by participating in hepcidin regulation via pathways that depend on AhR and oxidative stress.
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- 2018
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9. Methanol extraction fraction from Citrus Sudachi peel exerts lipid reducing effects in cultured cells.
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Xu W, Miyamoto L, Aihara H, Yamaoka T, Tanaka N, Tsuchihashi Y, Ikeda Y, Tamaki T, Kashiwada Y, and Tsuchiya K
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- AMP-Activated Protein Kinases metabolism, Animals, Cell Line, Humans, Hypolipidemic Agents isolation & purification, Hypolipidemic Agents pharmacology, Methanol, Mice, Models, Biological, PPAR alpha metabolism, Phytotherapy, Plant Extracts isolation & purification, Signal Transduction drug effects, Sirtuin 1 metabolism, Triglycerides metabolism, Citrus chemistry, Lipid Metabolism drug effects, Plant Extracts pharmacology
- Abstract
Ectopic fat accumulation is associated with insulin resistance and type 2 diabetes mellitus. Citrus sudachi is an evergreen tree that is found mainly in Tokushima Prefecture in Japan. Previously, it was demonstrated that Citrus sudachi could inhibit the rising trend of blood glucose and fatty acid in human subjects. In the current study, we illustrated the function of methanol extracts from sudachi peel and investigated the mechanism of this effect. We got the five kinds of methanol extracts by using diaion HP-20, and those were named by hydrophobicity from M-F1 to M-F5. Among the 5 kinds of sudachi methanol extracts, only M-F4 significantly decreased the intracellular triglyceride of C2C12 cells. It augmented the AMPK activity and increased the transcription of PPARα and its downstream targets CPT-1b and UCP2. In conclusion, M-F4 improved the lipid metabolism possibly through AMPK, PPARα and their downstream targets like CPT-1b and UCP2. Furthermore, this extract may be useful for preventing obesity and diabetes related diseases. J. Med. Invest. 65:225-230, August, 2018.
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- 2018
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10. Mechanisms of the pH- and Oxygen-Dependent Oxidation Activities of Artesunate.
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Tsuda K, Miyamoto L, Hamano S, Morimoto Y, Kangawa Y, Fukue C, Kagawa Y, Horinouchi Y, Xu W, Ikeda Y, Tamaki T, and Tsuchiya K
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- Antimalarials pharmacology, Antipyrine analogs & derivatives, Antipyrine pharmacology, Artesunate, Cell Survival drug effects, Edaravone, Free Radical Scavengers pharmacology, Hep G2 Cells, Humans, Hydrogen-Ion Concentration, Oxidation-Reduction, Antineoplastic Agents pharmacology, Artemisinins pharmacology, Iron pharmacology, Oxygen pharmacology, Reactive Oxygen Species metabolism
- Abstract
Artemisinin was discovered in 1971 as a constituent of the wormwood genus plant (Artemisia annua). This plant has been used as an herbal medicine to treat malaria since ancient times. The compound artemisinin has a sesquiterpene lactone bearing a peroxide group that offers its biological activity. In addition to anti-malarial activity, artemisinin derivatives have been reported to exert antitumor activity in cancer cells, and have attracted attention as potential anti-cancer drugs. Mechanisms that might explain the antitumor activities of artemisinin derivatives reportedly induction of apoptosis, angiogenesis inhibitory effects, inhibition of hypoxia-inducible factor-1α (HIF-1α) activation, and direct DNA injury. Reactive oxygen species (ROS) generation is involved in many cases. However, little is known about the mechanism of ROS formation from artemisinin derivatives and what types of ROS are produced. Therefore, we investigated the iron-induced ROS formation mechanism by using artesunate, a water-soluble artemisinin derivative, which is thought to be the underlying mechanism involved in artesunate-mediated cell death. The ROS generated by the coexistence of iron(II), artesunate, and molecular oxygen was a hydroxyl radical or hydroxyl radical-like ROS. Artesunate can reduce iron(III) to iron(II), which enables generation of ROS irrespective of the iron valence. We found that reduction from iron(III) to iron(II) was activated in the acidic rather than the neutral region and was proportional to the hydrogen ion concentration.
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- 2018
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11. Molecular Pathogenesis of Familial Wolff-Parkinson-White Syndrome.
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Miyamoto L
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- AMP-Activated Protein Kinases genetics, Animals, Humans, Wolff-Parkinson-White Syndrome genetics, AMP-Activated Protein Kinases physiology, Glycogen metabolism, Mutation, Myocardium metabolism, Wolff-Parkinson-White Syndrome etiology
- Abstract
Familial Wolff-Parkinson-White (WPW) syndrome is an autosomal dominant inherited disease and consists of a small percentage of WPW syndrome which exhibits ventricular pre-excitation by development of accessory atrioventricular pathway. A series of mutations in PRKAG2 gene encoding gamma2 subunit of 5'AMP-activated protein kinase (AMPK) has been identified as the cause of familial WPW syndrome. AMPK is one of the most important metabolic regulators of carbohydrates and lipids in many types of tissues including cardiac and skeletal muscles. Patients and animals with the mutation in PRKAG2 gene exhibit aberrant atrioventricular conduction associated with cardiac glycogen overload. Recent studies have revealed "novel" significance of canonical pathways leading to glycogen synthesis and provided us profound insights into molecular mechanism of the regulation of glycogen metabolism by AMPK. This review focuses on the molecular basis of the pathogenesis of cardiac abnormality due to PRKAG2 mutation and will provide current overviews of the mechanism of glycogen regulation by AMPK. J. Med. Invest. 65:1-8, February, 2018.
- Published
- 2018
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12. Dietary iron restriction alleviates renal tubulointerstitial injury induced by protein overload in mice.
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Ikeda Y, Horinouchi Y, Hamano H, Hirayama T, Kishi S, Izawa-Ishizawa Y, Imanishi M, Zamami Y, Takechi K, Miyamoto L, Ishizawa K, Aihara KI, Nagasawa H, Tsuchiya K, and Tamaki T
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- Acute Kidney Injury diet therapy, Acute Kidney Injury pathology, Animals, Biomarkers, Cytokines metabolism, Disease Models, Animal, Immunohistochemistry, Inflammasomes metabolism, Inflammation Mediators metabolism, Kidney Function Tests, Male, Mice, NADPH Oxidases metabolism, NLR Family, Pyrin Domain-Containing 3 Protein metabolism, Nephritis, Interstitial diet therapy, Nephritis, Interstitial pathology, Oxidative Stress, Serum Albumin, Bovine administration & dosage, Serum Albumin, Bovine adverse effects, Acute Kidney Injury etiology, Acute Kidney Injury metabolism, Iron metabolism, Iron, Dietary, Nephritis, Interstitial etiology, Nephritis, Interstitial metabolism, Proteinuria complications, Proteinuria metabolism
- Abstract
Increased proteinuria causes tubulointerstitial injury due to inflammation in chronic kidney disease (CKD). Iron restriction exhibits protective effects against renal dysfunction; however, its effects against protein overload-induced tubulointerstitial damage remain unclear. Here, we investigated dietary iron restriction effect on tubulointerstitial damage in mice with protein-overload tubulointerstitial injury. Renal tubulointerstitial injury in animal model was induced by intraperitoneal injection of an overdose of bovine serum albumin (BSA). We divided mice into three groups: normal saline + normal diet (ND), BSA + ND, and BSA + iron-restricted diet (IRD). BSA overload induced renal tubulointerstitial injury in the ND mice, which was ameliorated in the IRD mice. Inflammatory cytokines and extracellular matrix mRNA expression was upregulated in BSA + ND mice kidneys and was inhibited by IRD. BSA-induced increase in renal superoxide production, NADPH oxidase activity, and p22
phox expression was diminished in the IRD mice. IRD suppression increased BSA-induced renal macrophage infiltration. Moreover, BSA mice exhibited nucleotide-binding oligomerisation domain-like receptor pyrin domain-containing protein (NLRP) inflammasome activation, which was inhibited by IRD. Ferrous iron increased in kidneys with BSA overload and was inhibited by IRD. Thus, iron restriction inhibited oxidative stress and inflammatory changes, contributing to the protective effect against BSA overload-induced tubulointerstitial injury.- Published
- 2017
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13. Iron suppresses erythropoietin expression via oxidative stress-dependent hypoxia-inducible factor-2 alpha inactivation.
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Oshima K, Ikeda Y, Horinouchi Y, Watanabe H, Hamano H, Kihira Y, Kishi S, Izawa-Ishizawa Y, Miyamoto L, Hirayama T, Nagasawa H, Ishizawa K, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Erythropoietin analysis, Ferric Compounds pharmacology, Ferric Oxide, Saccharated, Fibroblasts, Glucaric Acid pharmacology, Hep G2 Cells, Humans, Kidney drug effects, Kidney metabolism, Mice, Up-Regulation drug effects, Ureteral Obstruction, Basic Helix-Loop-Helix Transcription Factors metabolism, Erythropoietin metabolism, Iron pharmacology, Oxidative Stress drug effects
- Abstract
Renal anemia is a major complication in chronic kidney disease (CKD). Iron supplementation, as well as erythropoiesis-stimulating agents, are widely used for treatment of renal anemia. However, excess iron causes oxidative stress via the Fenton reaction, and iron supplementation might damage remnant renal function including erythropoietin (EPO) production in CKD. EPO gene expression was suppressed in mice following direct iron treatment. Hypoxia-inducible factor-2 alpha (HIF-2α), a positive regulator of the EPO gene, was also diminished in the kidney of mice following iron treatment. Anemia-induced increase in renal EPO and HIF-2α expression was inhibited by iron treatment. In in vitro experiments using EPO-producing HepG2 cells, iron stimulation reduced the expression of the EPO gene, as well as HIF-2α. Moreover, iron treatment augmented oxidative stress, and iron-induced reduction of EPO and HIF-2α expression was restored by tempol, an antioxidant compound. HIF-2α interaction with the Epo promoter was inhibited by iron treatment, and was restored by tempol. These findings suggested that iron supplementation reduced EPO gene expression via an oxidative stress-HIF-2α-dependent signaling pathway.
- Published
- 2017
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14. Nitrite Activates 5'AMP-Activated Protein Kinase-Endothelial Nitric Oxide Synthase Pathway in Human Glomerular Endothelial Cells.
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Miyamoto L, Yamane M, Tomida Y, Kono M, Yamaoka T, Kawasaki A, Hatano A, Tsuda K, Xu W, Ikeda Y, Tamaki T, and Tsuchiya K
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- Cell Line, Endothelial Cells drug effects, Humans, Kidney Glomerulus cytology, Kidney Glomerulus drug effects, Nitric Oxide metabolism, Phosphorylation, AMP-Activated Protein Kinases metabolism, Endothelial Cells metabolism, Nitric Oxide Synthase Type III metabolism, Nitrites pharmacology, Protective Agents pharmacology, Signal Transduction drug effects
- Abstract
Recent studies have shown that orally supplied nitrates, which substantially exist in our daily diets, are reduced into nitrites and become significant sources of nitric oxide (NO) especially in hypoxic tissues. However, physiological significance of nitrites in normal tissues has not been elucidated though our serum concentrations of nitrites reach as high as micromolar levels. We investigated effects of nitrite on endothelial NO synthase (eNOS) using human glomerular endothelial cells to reveal potential glomerular-protective actions of nitrites with its underlying molecular mechanism. Here we demonstrate that nitrite stimulation evokes eNOS activation which is dependent on 5'AMP-activated protein kinase (AMPK) activation in accordance with ATP reduction. Thus, nitrites should facilitate AMPK-eNOS pathway in an energy level-dependent manner in endothelial cells. The activation of AMPK-eNOS signals is suggested to be involved in vascular and renal protective effects of nitrites and nitrates. Nitrites may harbor beneficial effects on metabolic regulations as AMPK activators.
- Published
- 2017
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15. Antitumor effect of novel anti-podoplanin antibody NZ-12 against malignant pleural mesothelioma in an orthotopic xenograft model.
- Author
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Abe S, Kaneko MK, Tsuchihashi Y, Izumi T, Ogasawara S, Okada N, Sato C, Tobiume M, Otsuka K, Miyamoto L, Tsuchiya K, Kawazoe K, Kato Y, and Nishioka Y
- Subjects
- Animals, Antibody-Dependent Cell Cytotoxicity immunology, Cell Line, Tumor, Complement System Proteins immunology, Cytotoxicity, Immunologic, Disease Models, Animal, Drug Therapy, Combination, Humans, Immunotherapy, Lung Neoplasms drug therapy, Lung Neoplasms pathology, Male, Mesothelioma drug therapy, Mesothelioma pathology, Mesothelioma, Malignant, Mice, Pemetrexed pharmacology, Pleural Neoplasms drug therapy, Pleural Neoplasms pathology, Rats, Xenograft Model Antitumor Assays, Antibodies, Monoclonal pharmacology, Antineoplastic Agents pharmacology, Lung Neoplasms immunology, Lung Neoplasms metabolism, Membrane Glycoproteins antagonists & inhibitors, Mesothelioma immunology, Mesothelioma metabolism, Pleural Neoplasms immunology, Pleural Neoplasms metabolism
- Abstract
Podoplanin (aggrus) is highly expressed in several types of cancers, including malignant pleural mesothelioma (MPM). Previously, we developed a rat anti-human podoplanin mAb, NZ-1, and a rat-human chimeric anti-human podoplanin antibody, NZ-8, derived from NZ-1, which induced antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity against podoplanin-positive MPM cell lines. In this study, we showed the antitumor effect of NZ-1, NZ-8, and NZ-12, a novel rat-human chimeric anti-human podoplanin antibody derived from NZ-1, in an MPM orthotopic xenograft SCID mouse model. Treatment with NZ-1 and rat NK (CD161a(+) ) cells inhibited the growth of tumors and the production of pleural effusion in NCI-H290/PDPN or NCI-H226 orthotopic xenograft mouse models. NZ-8 and human natural killer (NK) (CD56(+) ) cells also inhibited tumor growth and pleural effusion in MPM orthotopic xenograft mice. Furthermore, NZ-12 induced potent ADCC mediated by human MNC, compared with either NZ-1 or NZ-8. Antitumor effects were observed following treatment with NZ-12 and human NK (CD56(+) ) cells in MPM orthotopic xenograft mice. In addition, combined immunotherapy using the ADCC activity of NZ-12 mediated by human NK (CD56(+) ) cells with pemetrexed, led to enhanced antitumor effects in MPM orthotopic xenograft mice. These results strongly suggest that combination therapy with podoplanin-targeting immunotherapy using both NZ-12 and pemetrexed might provide an efficacious therapeutic strategy for the treatment of MPM., (© 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
- Published
- 2016
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16. Can food factors provide Us with the similar beneficial effects of physical exercise?
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Miyamoto L
- Abstract
Metabolic diseases have got global health issues. Physical exercise as well as diet therapy is a potent strategy for fighting against the diseases. However, it is often difficult to continue to keep exercise regularly enough to take sufficient effect. Thus, good substitutes for the therapeutic exercise would be greatly beneficial. Recent studies have suggested that 5'AMP-activated protein kinase (AMPK) play important roles in the metabolic alterations by muscle contraction. The notion that AMPK mediates broad effects of physical exercise has been widely accepted, though it has been challenged by observations in some genetically AMPK-disrupted animals. We have demonstrated metabolome-wide significance of AMPK activation in contracting muscles. Thus, pharmacological activation of AMPK can be a promising way to obtain similar effects of the exercise. The relevance of AMPK will be introduced, and possible strategies for obtaining similar effects to the exercise from food factors will be discussed in the current review.
- Published
- 2016
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17. A long-term high-fat diet changes iron distribution in the body, increasing iron accumulation specifically in the mouse spleen.
- Author
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Yamano N, Ikeda Y, Sakama M, Izawa-Ishizawa Y, Kihira Y, Ishizawa K, Miyamoto L, Tomita S, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Cation Transport Proteins genetics, Cation Transport Proteins metabolism, Diabetes Mellitus, Type 2 etiology, Dietary Fats adverse effects, Duodenum metabolism, Ferritins blood, Hemoglobins metabolism, Intestinal Absorption genetics, Male, Mice, Inbred C57BL, Obesity etiology, Obesity metabolism, Oxidative Stress, RNA, Messenger metabolism, Diabetes Mellitus, Type 2 metabolism, Diet, High-Fat adverse effects, Dietary Fats administration & dosage, Iron metabolism, Iron, Dietary metabolism, Spleen metabolism
- Abstract
Although iron is an essential trace metal, its presence in excess causes oxidative stress in the human body. Recent studies have indicated that iron storage is a risk factor for type 2 diabetes mellitus. Dietary iron restriction or iron chelation ameliorates symptoms of type 2 diabetes in mouse models. However, whether iron content in the body changes with the development of diabetes is unknown. Here, we investigated the dynamics of iron accumulation and changes in iron absorption-related genes in mice that developed obesity and diabetes by consuming a high-fat diet (HFD-fed mice). HFD-fed mice (18-20 wk) were compared with control mice for hematologic features, serum ferritin levels, and iron contents in the gastrocnemius muscle, heart, epididymal fat, testis, liver, duodenum, and spleen. In addition, the spleen was examined histologically. Iron absorption-related gene expression in the liver and duodenum was also examined. Hemoglobin and serum ferritin levels were increased in HFD-fed mice. The HFD-fed mice showed iron accumulation in the spleen, but not in the heart or liver. Increased percentages of the splenic red pulp and macrophages were observed in HFD-fed mice and iron accumulation in the spleen was found mainly in the splenic red pulp. The HFD-fed mice also showed decreased iron content in the duodenum. The mRNA expression of divalent metal transporter-1 (DMT-1), an iron absorption-related gene, was elevated in the duodenum of HFD-fed mice. These results indicate that iron accumulation (specifically accumulation in the spleen) is enhanced by the development of type 2 diabetes induced by HFD.
- Published
- 2015
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18. Iron chelation by deferoxamine prevents renal interstitial fibrosis in mice with unilateral ureteral obstruction.
- Author
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Ikeda Y, Ozono I, Tajima S, Imao M, Horinouchi Y, Izawa-Ishizawa Y, Kihira Y, Miyamoto L, Ishizawa K, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Blotting, Western, Fibrosis, Immunohistochemistry, Kidney Diseases etiology, Mice, Signal Transduction drug effects, Transforming Growth Factor beta metabolism, Chelation Therapy methods, Deferoxamine pharmacology, Iron Chelating Agents pharmacology, Kidney Diseases pathology, Kidney Diseases prevention & control, Ureteral Obstruction complications
- Abstract
Renal fibrosis plays an important role in the onset and progression of chronic kidney diseases (CKD). Although several mechanisms underlying renal fibrosis and candidate drugs for its treatment have been identified, the effect of iron chelator on renal fibrosis remains unclear. In the present study, we examined the effect of an iron chelator, deferoxamine (DFO), on renal fibrosis in mice with surgically induced unilateral ureter obstruction (UUO). Mice were divided into 4 groups: UUO with vehicle, UUO with DFO, sham with vehicle, and sham with DFO. One week after surgery, augmented renal tubulointerstitial fibrosis and the expression of collagen I, III, and IV increased in mice with UUO; these changes were suppressed by DFO treatment. Similarly, UUO-induced macrophage infiltration of renal interstitial tubules was reduced in UUO mice treated with DFO. UUO-induced expression of inflammatory cytokines and extracellular matrix proteins was abrogated by DFO treatment. DFO inhibited the activation of the transforming growth factor-β1 (TGF-β1)-Smad3 pathway in UUO mice. UUO-induced NADPH oxidase activity and p22(phox) expression were attenuated by DFO. In the kidneys of UUO mice, divalent metal transporter 1, ferroportin, and ferritin expression was higher and transferrin receptor expression was lower than in sham-operated mice. Increased renal iron content was observed in UUO mice, which was reduced by DFO treatment. These results suggest that iron reduction by DFO prevents renal tubulointerstitial fibrosis by regulating TGF-β-Smad signaling, oxidative stress, and inflammatory responses.
- Published
- 2014
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19. Nitrosonifedipine ameliorates the progression of type 2 diabetic nephropathy by exerting antioxidative effects.
- Author
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Ishizawa K, Izawa-Ishizawa Y, Yamano N, Urushihara M, Sakurada T, Imanishi M, Fujii S, Nuno A, Miyamoto L, Kihira Y, Ikeda Y, Kagami S, Kobori H, Tsuchiya K, and Tamaki T
- Subjects
- Animals, Cell Line, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Nifedipine therapeutic use, Nitric Oxide Synthase Type III genetics, Nitric Oxide Synthase Type III metabolism, Oxidative Stress drug effects, Antioxidants therapeutic use, Diabetic Nephropathies drug therapy, Diabetic Nephropathies metabolism, Nifedipine analogs & derivatives, Nitroso Compounds therapeutic use
- Abstract
Diabetic nephropathy (DN) is the major cause of end-stage renal failure. Oxidative stress is implicated in the pathogenesis of DN. Nitrosonifedipine (NO-NIF) is a weak calcium channel blocker that is converted from nifedipine under light exposure. Recently, we reported that NO-NIF has potential as a novel antioxidant with radical scavenging abilities and has the capacity to treat vascular dysfunction by exerting an endothelial protective effect. In the present study, we extended these findings by evaluating the efficacy of NO-NIF against DN and by clarifying the mechanisms of its antioxidative effect. In a model of type 2 DN (established in KKAy mice), NO-NIF administration reduced albuminuria and proteinuria as well as glomerular expansion without affecting glucose metabolism or systolic blood pressure. NO-NIF also suppressed renal and systemic oxidative stress and decreased the expression of intercellular adhesion molecule (ICAM)-1, a marker of endothelial cell injury, in the glomeruli of the KKAy mice. Similarly, NO-NIF reduced albuminuria, oxidative stress, and ICAM-1 expression in endothelial nitric oxide synthase (eNOS) knockout mice. Moreover, NO-NIF suppressed urinary angiotensinogen (AGT) excretion and intrarenal AGT protein expression in proximal tubular cells in the KKAy mice. On the other hand, hyperglycemia-induced mitochondrial superoxide production was not attenuated by NO-NIF in cultured endothelial cells. These findings suggest that NO-NIF prevents the progression of type 2 DN associated with endothelial dysfunction through selective antioxidative effects.
- Published
- 2014
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20. AICAR stimulation metabolome widely mimics electrical contraction in isolated rat epitrochlearis muscle.
- Author
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Miyamoto L, Egawa T, Oshima R, Kurogi E, Tomida Y, Tsuchiya K, and Hayashi T
- Subjects
- AMP-Activated Protein Kinases metabolism, Aminoimidazole Carboxamide pharmacology, Animals, Cell Line, Electric Stimulation, Factor Analysis, Statistical, Glucose metabolism, Glutathione metabolism, Male, Metabolome, Muscle Contraction physiology, Muscle, Skeletal physiology, Principal Component Analysis, Random Allocation, Rats, Rats, Sprague-Dawley, Transcriptome, Aminoimidazole Carboxamide analogs & derivatives, Hypoglycemic Agents pharmacology, Muscle Contraction drug effects, Muscle, Skeletal drug effects, Ribonucleotides pharmacology
- Abstract
Physical exercise has potent therapeutic and preventive effects against metabolic disorders. A number of studies have suggested that 5'-AMP-activated protein kinase (AMPK) plays a pivotal role in regulating carbohydrate and lipid metabolism in contracting skeletal muscles, while several genetically manipulated animal models revealed the significance of AMPK-independent pathways. To elucidate significance of AMPK and AMPK-independent signals in contracting skeletal muscles, we conducted a metabolomic analysis that compared the metabolic effects of 5-aminoimidazole-4-carboxamide-1-β-D-ribonucleoside (AICAR) stimulation with the electrical contraction ex vivo in isolated rat epitrochlearis muscles, in which both α1- and α2-isoforms of AMPK and glucose uptake were equally activated. The metabolomic analysis using capillary electrophoresis time-of-flight mass spectrometry detected 184 peaks and successfully annotated 132 small molecules. AICAR stimulation exhibited high similarity to the electrical contraction in overall metabolites. Principal component analysis (PCA) demonstrated that the major principal component characterized common effects whereas the minor principal component distinguished the difference. PCA and a factor analysis suggested a substantial change in redox status as a result of AMPK activation. We also found a decrease in reduced glutathione levels in both AICAR-stimulated and contracting muscles. The muscle contraction-evoked influences related to the metabolism of amino acids, in particular, aspartate, alanine, or lysine, are supposed to be independent of AMPK activation. Our results substantiate the significance of AMPK activation in contracting skeletal muscles and provide novel evidence that AICAR stimulation closely mimics the metabolomic changes in the contracting skeletal muscles.
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- 2013
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21. Leptin activates hepatic 5'-AMP-activated protein kinase through sympathetic nervous system and α1-adrenergic receptor: a potential mechanism for improvement of fatty liver in lipodystrophy by leptin.
- Author
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Miyamoto L, Ebihara K, Kusakabe T, Aotani D, Yamamoto-Kataoka S, Sakai T, Aizawa-Abe M, Yamamoto Y, Fujikura J, Hayashi T, Hosoda K, and Nakao K
- Subjects
- AMP-Activated Protein Kinases genetics, Animals, Cells, Cultured, Fatty Liver drug therapy, Fatty Liver genetics, Fatty Liver metabolism, Female, Hepatocytes enzymology, Hepatocytes metabolism, Humans, Leptin therapeutic use, Lipodystrophy drug therapy, Lipodystrophy genetics, Lipodystrophy metabolism, Male, Mice, Mice, Inbred C57BL, Muscle, Skeletal enzymology, Muscle, Skeletal metabolism, Rats, Rats, Wistar, Receptors, Adrenergic, alpha-1 genetics, Sympathetic Nervous System metabolism, AMP-Activated Protein Kinases metabolism, Fatty Liver enzymology, Leptin metabolism, Lipodystrophy enzymology, Receptors, Adrenergic, alpha-1 metabolism, Sympathetic Nervous System enzymology
- Abstract
Background: AMPK activation promotes glucose and lipid metabolism., Results: Hepatic AMPK activities were decreased in fatty liver from lipodystrophic mice, and leptin activated the hepatic AMPK via the α-adrenergic effect., Conclusion: Leptin improved the fatty liver possibly by activating hepatic AMPK through the central and sympathetic nervous systems., Significance: Hepatic AMPK plays significant roles in the pathophysiology of lipodystrophy and metabolic action of leptin. Leptin is an adipocyte-derived hormone that regulates energy homeostasis. Leptin treatment strikingly ameliorates metabolic disorders of lipodystrophy, which exhibits ectopic fat accumulation and severe insulin-resistant diabetes due to a paucity of adipose tissue. Although leptin is shown to activate 5'-AMP-activated protein kinase (AMPK) in the skeletal muscle, the effect of leptin in the liver is still unclear. We investigated the effect of leptin on hepatic AMPK and its pathophysiological relevance in A-ZIP/F-1 mice, a model of generalized lipodystrophy. Here, we demonstrated that leptin activates hepatic AMPK through the central nervous system and α-adrenergic sympathetic nerves. AMPK activities were decreased in the fatty liver of A-ZIP/F-1 mice, and leptin administration increased AMPK activities in the liver as well as in skeletal muscle with significant reduction in triglyceride content. Activation of hepatic AMPK with A769662 also led to a decrease in hepatic triglyceride content and blood glucose levels in A-ZIP/F-1 mice. These results indicate that the down-regulation of hepatic AMPK activities plays a pathophysiological role in the metabolic disturbances of lipodystrophy, and the hepatic AMPK activation is involved in the therapeutic effects of leptin.
- Published
- 2012
- Full Text
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22. Amylin improves the effect of leptin on insulin sensitivity in leptin-resistant diet-induced obese mice.
- Author
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Kusakabe T, Ebihara K, Sakai T, Miyamoto L, Aotani D, Yamamoto Y, Yamamoto-Kataoka S, Aizawa-Abe M, Fujikura J, Hosoda K, and Nakao K
- Subjects
- Animals, Anti-Obesity Agents administration & dosage, Delayed-Action Preparations administration & dosage, Delayed-Action Preparations therapeutic use, Diabetes Mellitus, Type 2 etiology, Diet, High-Fat adverse effects, Drug Resistance, Drug Therapy, Combination, Energy Intake drug effects, Energy Metabolism drug effects, Hypoglycemic Agents administration & dosage, Insulin blood, Islet Amyloid Polypeptide administration & dosage, Leptin administration & dosage, Liver drug effects, Liver metabolism, Male, Mice, Mice, Inbred C57BL, Muscle, Skeletal drug effects, Muscle, Skeletal metabolism, Obesity etiology, Obesity metabolism, Obesity physiopathology, Triglycerides metabolism, Weight Loss drug effects, Anti-Obesity Agents therapeutic use, Diabetes Mellitus, Type 2 prevention & control, Hypoglycemic Agents therapeutic use, Insulin Resistance, Islet Amyloid Polypeptide therapeutic use, Leptin therapeutic use, Obesity drug therapy
- Abstract
Leptin enhances insulin sensitivity in addition to reducing food intake and body weight. Recently, amylin, a pancreatic β-cell-derived hormone, was shown to restore a weight-reducing effect of leptin in leptin-resistant diet-induced obesity. However, whether amylin improves the effect of leptin on insulin sensitivity in diet-induced obesity is unclear. Diet-induced obese (DIO) mice were infused with either saline (S), leptin (L; 500 μg·kg⁻¹·day⁻¹), amylin (A; 100 μg·kg⁻¹·day⁻¹), or leptin plus amylin (L/A) for 14 days using osmotic minipumps. Food intake, body weight, metabolic parameters, tissue triglyceride content, and AMP-activated protein kinase (AMPK) activity were examined. Pair-feeding and weight-matched calorie restriction experiments were performed to assess the influence of food intake and body weight reduction. Continuous L/A coadministration significantly reduced food intake, increased energy expenditure, and reduced body weight, whereas administration of L or A alone had no effects. L/A coadministration did not affect blood glucose levels during ad libitum feeding but decreased plasma insulin levels significantly (by 48%), suggesting the enhancement of insulin sensitivity. Insulin tolerance test actually showed the increased effect of insulin in L/A-treated mice. In addition, L/A coadministration significantly decreased tissue triglyceride content and increased AMPKα2 activity in skeletal muscle (by 67%). L/A coadministration enhanced insulin sensitivity more than pair-feeding and weight-matched calorie restriction. In conclusion, this study demonstrates the beneficial effect of L/A coadministration on glucose and lipid metabolism in DIO mice, indicating the possible clinical usefulness of L/A coadministration as a new antidiabetic treatment in obesity-associated diabetes.
- Published
- 2012
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23. Cytotoxicity evaluation of symmetrically branched glycerol trimer in human hepatocellular carcinoma HepG2 cells.
- Author
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Miyamoto L, Watanabe M, Kono M, Matsushita T, Hattori H, Ishizawa K, Nemoto H, and Tsuchiya K
- Subjects
- Carcinoma, Hepatocellular, Cell Survival drug effects, Fenofibrate chemistry, Glycerol analogs & derivatives, Glycerol chemistry, Hep G2 Cells, Humans, Hydrophobic and Hydrophilic Interactions, Liver Neoplasms, Solubility, Glycerol toxicity
- Abstract
An appropriate balance between lipophilicity and hydrophilicity is necessary for pharmaceuticals to achieve fine Absorption, Distribution, Metabolism and Excretion (ADME) properties including absorption and distribution, in particular. We have designed and proposed symmetrically branched oligoglycerols (BGL) as an alternative approach to improve the lipophilic-hydrophilic balance. We have previously shown that stability in circulation and water-solubility of such molecules as proteins, liposomes and hydrophobic compounds are much improved by conjugation to BGL. Albeit these successful applications of BGL, little was known whether BGL could be used in safety. Thus we conducted evaluation of the cytotoxicity of a representative BGL, symmetrically branched glycerol trimer (BGL003) in the cultured cells to clarify its biological safeness. Here we demonstrate that water-solubility of an extremely hydrophobic agent, fenofibrate was more than 2,000-fold improved just by conjugated with BGL003. BGL003 did not exhibit any significant cytotoxicity in human hepatocarcinoma HepG2 cells. Thus BGL003 should be safe and suitable strategy to endow hydrophobic molecules with much hydrophilicity.
- Published
- 2012
- Full Text
- View/download PDF
24. Acute oral toxicity evaluation of symmetrically branched glycerol trimer in ddY mice.
- Author
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Miyamoto L, Watanabe M, Tomida Y, Kono M, Fujii S, Matsushita T, Hattori H, Ishizawa K, Nemoto H, and Tsuchiya K
- Subjects
- Administration, Oral, Animals, Female, Glycerol administration & dosage, Humans, Lethal Dose 50, Male, Mice, Mice, Inbred Strains, Molecular Conformation, Polymers, Solubility, Drug Design, Glycerol chemistry, Glycerol toxicity, Hydrophobic and Hydrophilic Interactions
- Abstract
Lipophilic-hydrophilic balance is a quite important determinant of pharmacokinetic properties of pharmaceuticals. Thus it is a key step to successfully manage lipophilic-hydrophilic balance in drug design. We have designed unique modular molecules, symmetrically branched oligoglycerols (BGL) as an alternative means to endow hydrophobic molecules with much hydrophilicity. We have succeeded in improving the water-solubility of several hydrophobic medicinal small molecules and thermal stability of artificial protein by covalent conjugation to BGL. We have also demonstrated that a representative BGL, symmetrically branched glycerol trimer (BGL003) does not exhibit significant cytotoxicity against human hepatocarcinoma HepG2 cells. However, there have been no reports suggesting whether BGL could be used in safety in vivo. Therefore, evaluation of acute oral toxicity of BGL003 in healthy mice was conducted. Here we demonstrate that an oral administration of BGL003 did not exhibit acute lethal toxicity up to 3,000 mg/kg. Body weight, food intake, blood glucose levels and weights of tissues were not affected by a short-term repetitive administration of increasing doses of BGL003. Biochemical indications related to hepatic disorders and tissue damage were unchanged, either. A single administration study revealed that 50% lethal dose of BGL003 should be more than 2,000 mg/kg. BGL003 will be safe and suitable approach to improve hydrophilicity of hydrophobic compounds.
- Published
- 2012
- Full Text
- View/download PDF
25. Effect of acute activation of 5'-AMP-activated protein kinase on glycogen regulation in isolated rat skeletal muscle.
- Author
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Miyamoto L, Toyoda T, Hayashi T, Yonemitsu S, Nakano M, Tanaka S, Ebihara K, Masuzaki H, Hosoda K, Ogawa Y, Inoue G, Fushiki T, and Nakao K
- Subjects
- AMP-Activated Protein Kinases, Aminoimidazole Carboxamide analogs & derivatives, Animals, Dose-Response Relationship, Drug, Enzyme Activation, Glucose metabolism, Glycogen Phosphorylase metabolism, Glycogen Synthase metabolism, Glycolysis physiology, In Vitro Techniques, Insulin physiology, Lactic Acid metabolism, Male, Rats, Rats, Sprague-Dawley, Ribonucleosides, Ribonucleotides, Time Factors, Adenosine Triphosphate metabolism, Glycogen metabolism, Multienzyme Complexes metabolism, Muscle Contraction physiology, Muscle, Skeletal metabolism, Protein Serine-Threonine Kinases metabolism
- Abstract
5'-AMP-activated protein kinase (AMPK) has been implicated in glycogen metabolism in skeletal muscle. However, the physiological relevance of increased AMPK activity during exercise has not been fully clarified. This study was performed to determine the direct effects of acute AMPK activation on muscle glycogen regulation. For this purpose, we used an isolated rat muscle preparation and pharmacologically activated AMPK with 5-aminoimidazole-4-carboxamide-1-beta-D-ribonucleoside (AICAR). Tetanic contraction in vitro markedly activated the alpha(1)- and alpha(2)-isoforms of AMPK, with a corresponding increase in the rate of 3-O-methylglucose uptake. Incubation with AICAR elicited similar enhancement of AMPK activity and 3-O-methylglucose uptake in rat epitrochlearis muscle. In contrast, whereas contraction stimulated glycogen synthase (GS), AICAR treatment decreased GS activity. Insulin-stimulated GS activity also decreased after AICAR treatment. Whereas contraction activated glycogen phosphorylase (GP), AICAR did not alter GP activity. The muscle glycogen content decreased in response to contraction but was unchanged by AICAR. Lactate release was markedly increased when muscles were stimulated with AICAR in buffer containing glucose, indicating that the glucose taken up into the muscle was catabolized via glycolysis. Our results suggest that AMPK does not mediate contraction-stimulated glycogen synthesis or glycogenolysis in skeletal muscle and also that acute AMPK activation leads to an increased glycolytic flux by antagonizing contraction-stimulated glycogen synthesis.
- Published
- 2007
- Full Text
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26. Possible involvement of the alpha1 isoform of 5'AMP-activated protein kinase in oxidative stress-stimulated glucose transport in skeletal muscle.
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Toyoda T, Hayashi T, Miyamoto L, Yonemitsu S, Nakano M, Tanaka S, Ebihara K, Masuzaki H, Hosoda K, Inoue G, Otaka A, Sato K, Fushiki T, and Nakao K
- Subjects
- AMP-Activated Protein Kinases, Animals, Dose-Response Relationship, Drug, In Vitro Techniques, Isoenzymes antagonists & inhibitors, Isoenzymes metabolism, Leucine pharmacology, Male, Muscle, Skeletal drug effects, Oxidative Stress drug effects, Rats, Rats, Sprague-Dawley, Acetylcysteine pharmacology, Glucose metabolism, Hydrogen Peroxide pharmacology, Leucine analogs & derivatives, Multienzyme Complexes metabolism, Muscle, Skeletal metabolism, Oxidative Stress physiology, Protein Serine-Threonine Kinases metabolism
- Abstract
Recent studies have suggested that 5'AMP-activated protein kinase (AMPK) is activated in response to metabolic stresses, such as contraction, hypoxia, and the inhibition of oxidative phosphorylation, which leads to insulin-independent glucose transport in skeletal muscle. In the present study, we hypothesized that acute oxidative stress increases the rate of glucose transport via an AMPK-mediated mechanism. When rat epitrochlearis muscles were isolated and incubated in vitro in Krebs buffer containing the oxidative agent H(2)O(2), AMPKalpha1 activity increased in a time- and dose-dependent manner, whereas AMPKalpha2 activity remained unchanged. The activation of AMPKalpha1 was associated with phosphorylation of AMPK Thr(172), suggesting that an upstream kinase is involved in the activation process. H(2)O(2)-induced AMPKalpha1 activation was blocked in the presence of the antioxidant N-acetyl-l-cysteine (NAC), and H(2)O(2) significantly increased the ratio of oxidized glutathione to glutathione (GSSG/GSH) concentrations, a sensitive marker of oxidative stress. H(2)O(2) did not cause an increase in the conventional parameters of AMPK activation, such as AMP and AMP/ATP. H(2)O(2) increased 3-O-methyl-d-glucose transport, and this increase was partially, but significantly, blocked in the presence of NAC. Results were similar when the muscles were incubated in a superoxide-generating system using hypoxanthine and xanthine oxidase. Taken together, our data suggest that acute oxidative stress activates AMPKalpha1 in skeletal muscle via an AMP-independent mechanism and leads to an increase in the rate of glucose transport, at least in part, via an AMPKalpha1-mediated mechanism.
- Published
- 2004
- Full Text
- View/download PDF
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