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Elucidating the anti-colitic mechanisms of rice bran supplementation via nutriomics
Aw, WanPing
2020
科学研究費補助金研究成果報告書 (2019. )10％米ぬかを摂取したマウスが大腸炎発症による体重減少を抑制した．腸内細菌叢解析を行った
結果，米ぬかの摂取によってClostridiaceaeの割合が高くなったことが明らかになった。無菌マウスへ米ぬかを摂取したマウスの糞便を経口投与して、大腸炎発症による体重減少を抑制した。米ぬかを摂取したマウスの便中のトリプトファン関連代謝物には有意義に変動した。
Our results suggest that colitic mice fed 10% RB supplemented diet had higher abundance ofClostridiaceae. Fecal microbiota transplantation from RB-fed mice suppressed colitis. This strongly
suggests that RB suppresses colitis through gut microbiota modulation.The fecal metabolome
profile was dynamically changed by RB intake. Enrichment analysis revealed that RB interventionenriched the tryptophan metabolism pathway. Our results revealed that RB improves colitis via gut
environment modulation.
研究種目 : 若手研究
研究期間 : 2018～2019
課題番号 : 18K17976
研究分野 : Nutriomics
Research Paper
https://koara.lib.keio.ac.jp/xoonips/modules/xoonips/detail.php?koara_id=KAKEN_18K17976seika

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３版

様式

Ｃ−１９、Ｆ−１９−１、Ｚ−１９ （共通）

科学研究費助成事業

研究成果報告書
令和２ 年

６ 月 １８ 日現在

機関番号： ３２６１２
研究種目： 若手研究
研究期間： 2018 〜 2019
課題番号： １８Ｋ１７９７６
研究課題名（和文）Elucidating the anti‑colitic mechanisms of Rice Bran supplementation via
nutriomics
研究課題名（英文）Elucidating the anti‑colitic mechanisms of Rice Bran supplementation via
nutriomics
研究代表者
Ａｗ

ＷａｎＰｉｎｇ（AW, WanPing）

慶應義塾大学・政策・メディア研究科（藤沢）・特任助教

研究者番号：２０７３６４４１
交付決定額（研究期間全体）：（直接経費）

2,600,000 円

研究成果の概要（和文）：10％米ぬかを摂取したマウスが大腸炎発症による体重減少を抑制した．腸内細菌叢解析を行った結果，米ぬかの摂取によってClostridiaceaeの割合が高くなったことが明らかになった。無菌マウス
へ米ぬかを摂取したマウスの糞便を経口投与して、大腸炎発症による体重減少を抑制した。米ぬかを摂取したマウスの便中のトリプトファン関連代謝物には有意義に変動した。

研究成果の学術的意義や社会的意義
We have provided scientific evidence that rice bran suppresses colitis and we could apply it torecommending IBD patients to consume unpolished brown rice instead of white rice. Our methodology inelucidating the anti‑colitic mechanisms of rice bran could also be applied to other food products.

研究成果の概要（英文）：Our results suggest that colitic mice fed 10% RB supplemented diet had
higher abundance of Clostridiaceae. Fecal microbiota transplantation from RB‑fed mice suppressedcolitis. This strongly suggests that RB suppresses colitis through gut microbiota modulation.Thefecal metabolome profile was dynamically changed by RB intake. Enrichment analysis revealed that RB
intervention enriched the tryptophan metabolism pathway. Our results revealed that RB improves
colitis via gut environment modulation.

研究分野： Nutriomics
キーワード： Rice bran

colitisgut microbiome

metabolome※科研費による研究は、研究者の自覚と責任において実施するものです。そのため、研究の実施や研究成果の公表等については、国の要請等に基づくものではなく、その研究成果に関する見解や責任は、研究者個人に帰属されます。

様式 Ｃ−１９、Ｆ−１９−１、Ｚ−１９（共通）１．研究開始当初の背景
In recent years, the prevention of life-style related disease such Type 2 Diabetes (T2D)(Nutr Diabetes., 5 (11), e185, 2015), metabolic disease (Br. J. Nutr., 112: 717-719, 2014) andIBD (Sci. Rep., 7, 43993, 2017) via nutritional intervention is popular and consumers are
becoming more conscious of their nutritional intake to maintain good health. RB that is containedin brown rice, a type of Japanese food that is commonly consumed, has reported colitis
suppression effects, but the molecular mechanisms are not yet understood (Scand. J.
Gastroenterol., 46, 40–52, 2011).
２．研究の目的
The purpose and “key scientific questions” that would be answered in this project are (i)elucidating the molecular mechanisms of colitis suppression by RB and its components
via nutriomics analysis. I am using an original approach: nutriomics strives to promote agreater understanding of the ways nutrition influences metabolic pathways and homeostaticcontrol and how this regulation is disturbed in the early stages of a diet-related disease (Semin.
Immunopathol., 37 (1), 5-16, 2015). The gastrointestinal tract is a complex ecosystem thatharbors various microorganisms which influence many aspects of health including inflammatory
bowel disease, cancer and obesity in adults. The intricate interaction among gut microbiotaderived metabolites, the gut microbiota itself and the host immune system is transmitted via alarge array of signaling pathways that extend beyond the immune system. These complex interrelationships come together to form a series of host-microbe metabolic axes. By integrating
metagenomic and metabolomics information on a systems biology-wide approach, I would bebetter able to understand this interplay between RB, gut microbiome, host metabolism and IBD.
Through the modelling of metabolic interactions between RB and microbiota, I foresee that
integrated omics-based understanding of the gut ecosystem is the new avenue, providing exciting
novel therapeutic approaches for personalized host health.

３．研究の方法
We performed 3 animal experiments (2 animal experiments using conventional mice and 1 animal
experiment using germ-free mice). The 1st animal experiment was to observe dose dependencyof RB on colitis suppression. Following a 1-week acclimatization period, mice were randomly
assigned to 4 different groups (n=5 in each group): Control: mice given the AIN-93G basal dietand tap water for 1 week, after which colitis was induced by administering 2.0% (w/v) DSScontaining water for 10 days; 5% RB, 10% RB and 20% RB-supplemented AIN-93G basal dietand tap water for 1 week, then 2.0% DSS-containing water for 10 days respectively. During theexperimental period, fecal samples from all of the mice were collected daily for 16S rRNA gene
analysis. In addition, during the DSS administration period, severity of colitis would be assessed.
On day 10 after DSS administration, all mice would be sacrificed.
The 2nd animal experiment was to investigate fat-soluble/water-soluble components of RB on
colitis suppression: Following a 1-week acclimatization period, mice were randomly assigned to6 different groups (n=5 in each group): Control: mice given the AIN-93G basal diet and tap waterfor 1 week, after which colitis was induced by administering 2.0% (w/v) DSS-containing water for
10 days; RB, RBN, RBG and RBD: 10% RB, γ-oryzanol and 10% defatted RB-supplemented AIN93G basal diet and tap water for 1 week, then 2.0% DSS-containing water for 10 days respectively.
During the experimental period, fecal samples from all of the mice were collected daily for 16S
rRNA gene analysis. In addition, during the DSS administration period, severity of colitis would
be assessed. On day 10 after DSS administration, all mice would be sacrificed.
The germ-free murine experiment was to investigate if the colitis suppression effects from RB was
solely dependent on gut microbiome modulation. Briefly, 8-week-old specific-pathogen-free male
C5BL6J donor mice (n=3 per diet group) were administered with AIN-93G with tap water (CON),
10% supplemented RB diet with tap water (10% RB). After 2 weeks of administration, fecal
samples were collected and pooled. 100 mg of fecal sample was resuspended in 1 ml of sterile
PBS. The solution was vigorously mixed for 10s using a benchtop vortex (Vortex-Genie 2,Scientific Industries, USA; speed 9), before centrifugation at 800g for 3 min. The supernatant thatwas prepared on the same day of transplantation within 10 min before oral gavage to preventchanges in microbiota composition was collected and was administered into recipient mice daily(350 uL for each mouse) by oral gavage. Recipient mice were randomly assigned to 2 groups andwere fed with AIN-93G basal diet and tap water for 10 days, then administered 2.0% DSScontaining water for 10 days. Additionally, recipient mice were administered supernatant of fresh
fecal suspension from CON and 10% RB to their respective recipient groups, respectively, by oral

gavage (350 uL for each mouse) daily.
For all animal experiments, we performed the following observations.
Colon observations Length of colon and presence of tumors in colon will be macroscopicallyinspected. For histopathology analysis, a representative sample from the mid-part of colon will beselected.
Collection of tissues for multi-omics evaluation Fecal samples will be collected on asindicated above for metabolite analysis. Intestinal contents will also be collected from the ileum
and colon at time of sacrifice for metabolomics (CE-TOF/MS, Agilent, available at Keio University)and metagenomic (Miseq System, Illumina, available at Keio University) analyses.
Evaluation of omics data for integrated omics analysis One of the core goals of nutritional
research is to understand how diet can influence metabolic regulations and how scientists in thisfield can use this diet to improve health and well-being. I will integrate multiple omics platforms
and finally to systems biology by using software like Ingeunity Pathway Analysis (IPA, Qiagen)and MetaCore (Thomson Reuters) to have an overview of integrated omics data to visuallycompare the responses of multiple metabolic pathways simultaneously. The
pathways/genes/proteins/metabolites are activated or repressed among the multiple metabolic
pathways can be visually and intuitively found. Gut microbiome will be analysed using theQuantitative Insights Into Microbial Ecology (QIIME) platform.
４．研究成果In this study, we have shown that 5%. 10% and 20% RB fed mice have reported colitis
suppression (A). By using a time-course microbiome and metabolome approach, I aimed toelucidate the dynamics of gut environment, and identified RB-related gut environmental
alterations. 20 families, especially Alcaligenaceae and Lactobacillaceae showed significant
increases and Enterobacteriaceae was significantly decreased in 10% RB-fed mice (D).
Clostridiaceae had significantly higher abundances (J). 41 metabolites, especially N1,N12Diacetylspermine, Indole-3-ethanol, N1-Acetylspermidine, 5-Hydroxy-indoleacetate and shortchain fatty acids were significantly increased RB-fed mice (E,F). Metabolite set enrichment
analysis revealed that Vitamin B6 metabolism, tryptophan metabolism and ubiquinone
biosynthesis were significantly increased in RB-fed mice (G). Mice that underwent fecal
microbiota transplantation from donor mice fed RB had significantly improved body weight loss(I).

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