研究者詳細 - 伊藤　美佳子

Freeze-drying enables homogeneous and stable sample preparation for determination of fecal short-chain fatty acids 査読有り国際誌

Ueyama, J; Oda, M; Hirayama, M; Sugitate, K; Sakui, N; Hamada, R; Ito, M; Saito, I; Ohno, K

ANALYTICAL BIOCHEMISTRY 589 巻頁： 113508 - 113508 2020年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Analytical Biochemistry

Background: The analysis methods for fecal short-chain fatty acids (SCFAs) have evolved considerably. Recently, the role of SCFAs in gastrointestinal physiology and their association with intestinal microbiota and disease were reported. However, the intra-fecal variability and storage stability of SCFAs have not been extensively investigated. The aim of this study was to understand the limitations of the measurement of SCFAs in crude feces and develop a useful pre-examination procedure using the freeze-drying technique. Methods: SCFAs in crude feces, obtained from healthy volunteers, and freeze-dried feces were determined by derivatization with isobutyl chloroformate, followed by liquid–liquid extraction with hexane, and separation and analysis using gas chromatography–mass spectrometry. Results: Among the SCFAS, the maximum intra-fecal variability was observed for iso-butyrate (coefficient of variation of 37.7%), but the freeze-drying procedure reduced this variability (coefficient of variation of 7.9%). Similar improvements were also observed for other SCFAs. Furthermore, significant decreases in the SCFA amounts were observed with storage at 4 °C for 24 h. Conclusions: The freeze-drying procedure affords fecal SCFA stability, even with storage at room temperature for 3 d. The freeze-drying procedure allows reliable SCFA measurements without labour-intensive processes. Therefore, the freeze-drying procedure can be applied in basic, clinical, and epidemiological studies.

DOI： 10.1016/j.ab.2019.113508

Lewy小体病患者のメタメタ解析を用いた腸内細菌叢の関与の解明査読有り

平山正昭, 石田智大, 九鬼桃茄, 浜口知成, 伊藤美佳子, 上山純, 島村徹平, 阿部興, 勝野雅央, 大野欽司

臨床神経学 59 巻 ( Suppl. ) 頁： S301 - S301 2019年11月

パーキンソン病(PD)における腸内細菌査読有り

石田智大, 九鬼桃茄, 浜口知成, 上山純, 伊藤美佳子, 柏原健一, 前田哲也, 坪井義夫, 阿部興, 島村徹平, 森宙史, 黒川顕, 勝野雅央, 大野欽司, 平山正昭

パーキンソン病・運動障害疾患コングレスプログラム・抄録集 13回巻頁： 102 - 102 2019年7月

神経変性疾患における短鎖脂肪酸の役割査読有り

九鬼桃茄, 石田智大, 浜口知成, 伊藤美佳子, 木村郁夫, 長谷耕二, 橋本款, 大野欽司, 平山正昭

パーキンソン病・運動障害疾患コングレスプログラム・抄録集 13回巻頁： 102 - 102 2019年7月

Mianserin suppresses R-spondin 2-induced activation of Wnt/β-catenin signaling in chondrocytes and prevents cartilage degradation in a rat model of osteoarthritis 査読有り国際誌 Open Access

Okura, T; Ohkawara, B; Takegami, Y; Ito, M; Masuda, A; Seki, T; Ishiguro, N; Ohno, K

SCIENTIFIC REPORTS 9 巻 ( 1 ) 頁： 2808 - 2808 2019年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Scientific Reports

Aberrant activation of the Wnt/β-catenin signaling pathway promotes the progression of osteoarthritis (OA). We previously reported that R-spondin 2 (Rspo2), an activator of the Wnt/β-catenin signaling, facilitates differentiation of proliferating chondrocytes into hypertrophic chondrocytes by enhancing Wnt/β-catenin signaling in endochondral ossification. However, the role of Rspo2 in OA remains elusive. Here, we showed that the amounts of Rspo2 protein in synovial fluid were increased in OA patients. We searched for a preapproved drug that suppresses Rspo2-induced Wnt/β-catenin signaling in chondrogenic cells and reduces joint pathology in a rat model of OA. In Rspo2-treated ATDC5 cells, mianserin, a tetracyclic antidepressant, inhibited Wnt/β-catenin signaling, increased proteoglycan production, and upregulated chondrogenic marker genes. Mianserin suppressed Rspo2-induced accumulation of β-catenin and phosphorylation of Lrp6. We identified that mianserin blocked binding of Rspo2 to its receptor Lgr5. We also observed that intraarticular administration of mianserin suppressed β-catenin accumulation and prevented OA progression in a rat model of OA. We conclude that mianserin suppresses abnormally activated Wnt/β-catenin signaling in OA by inhibiting binding of Rspo2 to Lgr5.

DOI： 10.1038/s41598-019-39393-x

Quantification of hydrogen production by intestinal bacteria that are specifically dysregulated in Parkinson's disease 査読有り国際誌 Open Access

Suzuki, A; Ito, M; Hamaguchi, T; Mori, H; Takeda, Y; Baba, R; Watanabe, T; Kurokawa, K; Asakawa, S; Hirayama, M; Ohno, K

PLOS ONE 13 巻 ( 12 ) 頁： e0208313 2018年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PLoS ONE

Oral administration of hydrogen water ameliorates Parkinson’s disease (PD) in rats, mice, and humans. We previously reported that the number of putative hydrogen-producing bacteria in intestinal microbiota is low in PD compared to controls. We also reported that the amount of hydrogen produced by ingestion of lactulose is low in PD patients. The decreased hydrogen production by intestinal microbiota may be associated with the development and progression of PD. We measured the amount of hydrogen production using gas chromatography by seven bacterial strains, which represented seven major intestinal bacterial groups/ genera/species. Blautia coccoides and Clostridium leptum produced the largest amount of hydrogen. Escherichia coli and Bacteroides fragilis constituted the second group that produced hydrogen 34- to 93-fold lower than B. coccoides. Bifidobacterium pseudocatenulatum and Atopobium parvulum constituted the third group that produced hydrogen 559- to 2164-fold lower than B. coccoides. Lactobacillus casei produced no detectable hydrogen. Assuming that taxonomically neighboring strains have similar hydrogen production, we simulated hydrogen production using intestinal microbiota that we previously reported, and found that PD patients produce a 2.2-fold lower amount of intestinal hydrogen compared to controls. The lower amount of intestinal hydrogen production in PD was also simulated in cohorts of two other countries. The number of hydrogen-producing intestinal bacteria may be associated with the development and progression of PD. Further studies are required to prove its beneficial effect.

DOI： 10.1371/journal.pone.0208313

Inhalation of hydrogen gas elevates urinary 8-hydroxy-2'-deoxyguanine in Parkinson's disease. 査読有り国際誌 Open Access

Hirayama M, Ito M, Minato T, Yoritaka A, LeBaron TW, Ohno K

Medical gas research 8 巻 ( 4 ) 頁： 144 - 149 2018年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Medical Gas Research

Hyposmia is one of the earliest and the most common symptoms in Parkinson's disease (PD). The benefits of hydrogen water on motor deficits have been reported in animal PD models and PD patients, but the effects of hydrogen gas on PD patients have not been studied. We evaluated the effect of inhalation of hydrogen gas on olfactory function, non-motor symptoms, activities of daily living, and urinary 8-hydroxy-2′-deoxyguanine (8-OHdG) levels by a randomized, double-blinded, placebo-controlled, crossover trial with an 8-week washout period in 20 patients with PD. Patients inhaled either 1.2-1.4% hydrogen-air mixture or placebo for 10 minutes twice a day for 4 weeks. Inhalation of low dose hydrogen did not significantly influence the PD clinical parameters, but it did increase urinary 8-OHdG levels by 16%. This increase in 8-OHdG is markedly less than the over 300% increase in diabetes, and is more comparable to the increase after a bout of strenuous exercise. Although increased reactive oxygen species is often associated with toxicity and disease, they also play essential roles in mediating cytoprotective cellular adaptations in a process known as hormesis. Increases of oxidative stress by hydrogen have been previously reported, along with its ability to activate the Nrf2, NF-κB pathways, and heat shock responses. Although we did not observe any beneficial effect of hydrogen in our short trial, we propose that the increased 8-OHdG and other reported stress responses from hydrogen may indicate that its beneficial effects are partly or largely mediated by hormetic mechanisms. The study was approved by the ethics review committee of Nagoya University Graduate School of Medicine (approval number 2015-0295). The clinical trial was registered at the University Hospital Medical Information Network (identifier UMIN000019082).

DOI： 10.4103/2045-9912.248264

Differential effects of spinal motor neuron-derived and skeletal muscle-derived Rspo2 on acetylcholine receptor clustering at the neuromuscular junction 査読有り Open Access

Li, J; Ito, M; Ohkawara, B; Masuda, A; Ohno, K

SCIENTIFIC REPORTS 8 巻 ( 1 ) 頁： 13577 - - 2018年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Scientific Reports

We recently reported that R-spondin 2 (Rspo2), a secreted activator of Wnt/β-catenin signaling, promotes acetylcholine receptor (AChR) clustering and neuromuscular junction (NMJ) formation via its receptor, Lgr5. Rspo2 is expressed highly in spinal motor neurons (SMNs) and marginally in the skeletal muscle, but the origin of Rspo2 at the NMJ remains elusive. We rescued Rspo2-deficient (Rspo2−/−) mice by specifically expressing Rspo2 in the skeletal muscle and SMNs. SMN-specific Rspo2 mitigated or over-corrected abnormal features of the NMJs and AChR clusters observed in Rspo2−/− mice including (i) abnormal broadening of enlarged AChR clusters, (ii) three of six abnormal ultrastructural features, and (iii) abnormal expression of nine genes in SMNs and the diaphragm. In contrast, muscle-specific Rspo2 normalized all six abnormal ultrastructural features, but it had no effect on AChR clustering and NMJ formation at the light microscopy level or on abnormal gene expression in SMNs and the diaphragm. These results suggest that SMN-derived Rspo2 plays a major role in AChR clustering and NMJ formation in the postsynaptic region, and muscle-derived Rspo2 also plays a substantial role in juxtaposition of the active zones and synaptic folds.

DOI： 10.1038/s41598-018-31949-7

Randomized, Double-Blind, Multicenter Trial of Hydrogen Water for Parkinson's Disease 査読有り国際誌

Yoritaka, A; Ohtsuka, C; Maeda, T; Hirayama, M; Abe, T; Watanabe, H; Saiki, H; Oyama, G; Fukae, J; Shimo, Y; Hatano, T; Kawajiri, S; Okuma, Y; Machida, Y; Miwa, H; Suzuki, C; Kazama, A; Tomiyama, M; Kihara, T; Hirasawa, M; Shimura, H; Oda, E; Ito, M; Ohno, K; Hattori, N

MOVEMENT DISORDERS 33 巻 ( 9 ) 頁： 1505 - 1507 2018年9月

Protein-anchoring therapy to target extracellular matrix proteins to their physiological destinations 査読有り国際誌

Ito, M; Ohno, K

MATRIX BIOLOGY 68-69 巻 ( - ) 頁： 628 - 636 2018年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Matrix Biology

Endplate acetylcholinesterase (AChE) deficiency is a form of congenital myasthenic syndrome (CMS) caused by mutations in COLQ, which encodes collagen Q (ColQ). ColQ is an extracellular matrix (ECM) protein that anchors AChE to the synaptic basal lamina. Biglycan, encoded by BGN, is another ECM protein that binds to the dystrophin-associated protein complex (DAPC) on skeletal muscle, which links the actin cytoskeleton and ECM proteins to stabilize the sarcolemma during repeated muscle contractions. Upregulation of biglycan stabilizes the DPAC. Gene therapy can potentially ameliorate any disease that can be recapitulated in cultured cells. However, the difficulty of tissue-specific and developmental stage-specific regulated expression of transgenes, as well as the difficulty of introducing a transgene into all cells in a specific tissue, prevents us from successfully applying gene therapy to many human diseases. In contrast to intracellular proteins, an ECM protein is anchored to the target tissue via its specific binding affinity for protein(s) expressed on the cell surface within the target tissue. Exploiting this unique feature of ECM proteins, we developed protein-anchoring therapy in which a transgene product expressed even in remote tissues can be delivered and anchored to a target tissue using specific binding signals. We demonstrate the application of protein-anchoring therapy to two disease models. First, intravenous administration of adeno-associated virus (AAV) serotype 8-COLQ to Colq-deficient mice, resulting in specific anchoring of ectopically expressed ColQ-AChE at the NMJ, markedly improved motor functions, synaptic transmission, and the ultrastructure of the neuromuscular junction (NMJ). In the second example, Mdx mice, a model for Duchenne muscular dystrophy, were intravenously injected with AAV8-BGN. The treatment ameliorated motor deficits, mitigated muscle histopathologies, decreased plasma creatine kinase activities, and upregulated expression of utrophin and DAPC component proteins. We propose that protein-anchoring therapy could be applied to hereditary/acquired defects in ECM and secreted proteins, as well as therapeutic overexpression of such factors.

DOI： 10.1016/j.matbio.2018.02.014

Molecular hydrogen upregulates heat shock response and collagen biosynthesis, and downregulates cell cycles: meta-analyses of gene expression profiles 国際誌

Loss of Sfpq Causes Long-Gene Transcriptopathy in the Brain

Takeuchi Akihide, Iida Kei, Tsubota Toshiaki, Hosokawa Motoyasu, Denawa Masatsugu, Brown J. B., Ninomiya Kensuke, Ito Mikako, Kimura Hiroshi, Abe Takaya, Kiyonari Hiroshi, Ohno Kinji, Hagiwara Masatoshi

CELL REPORTS 23 巻 ( 5 ) 頁： 1326-1341 2018年5月

Nishiwaki, H; Ito, M; Negishi, S; Sobue, S; Ichihara, M; Ohno, K

FREE RADICAL RESEARCH 52 巻 ( 4 ) 頁： 434 - 445 2018年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Free Radical Research

Molecular hydrogen exerts its effect on multiple pathologies, including oxidative stress, inflammation, and apoptosis. However, its molecular mechanisms have not been fully elucidated. In order to explore the effects of molecular hydrogen, we meta-analysed gene expression profiles modulated by molecular hydrogen. We performed microarray analysis of the mouse liver with or without drinking hydrogen water. We also integrated two previously reported microarray datasets of the rat liver into meta-analyses. We used two categories of meta-analysis methods: the cross-platform method and the conventional meta-analysis method (Fisher’s method). For each method, hydrogen-modulated pathways were analysed by (i) the hypergeometric test (HGT) in the class of over-representation analysis (ORA), (ii) the gene set enrichment analysis (GSEA) in the class of functional class scoring (FCS), and (iii) the signalling pathway impact analysis (SPIA), pathway regulation score (PRS), and others in the class of pathway topology-based approach (PTA). Pathways in the collagen biosynthesis and the heat-shock response were up-regulated according to (a) HGT with the cross-platform method, (b) GSEA with the cross-platform method, and (c) PRS with the cross-platform method. Pathways in cell cycles were down-regulated according to (a) HGT with the cross-platform method, (b) GSEA with the cross-platform method, and (d) GSEA with the conventional meta-analysis method. Because the heat-shock response leads to up-regulation of collagen biosynthesis and a transient arrest of cell cycles, induction of the heat-shock response is likely to be a primary event induced by molecular hydrogen in the liver of wild-type rodents.

DOI： 10.1080/10715762.2018.1439166