Växtbaserade Proteiners påverkan på tarmhälsa Hos människa
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 59
- 试验地点
- 2
- 主要终点
- Changes in the faecal metabolites using targeted and untargeted metabolomics during the intervention
研究概览
简要总结
This project aims to create a methodological framework, including the discovery and validation of novel biomarker panels to decipher the impact of plant-based protein intake on health biomarkers. Moreover, the possibility of predicting biomarkers production will be tested by a colonic in vitro fermentation study using study participants' faecal samples.
详细描述
This study consists of a double-blind, randomised, 3-arm parallel trial, consisting of 2-wk baseline measurements, 4-wk intervention period (with weekly sample collection), and 2-wk after-intervention follow-up, with healthy subjects in each arm consuming different protein sources (from plant- or animal-based sources). Subjects that are eligible to take part in this study will be blinded and randomly allocated to consume an isolated pea protein (arm 1), animal protein (arm 2) or more complex protein pea protein powder (arm 3). The isolated plant and animal protein will be used to evaluate the specific effects of the proteins from the different sources. The more complex protein pea protein powder will be used as a more complex food matrix, containing other food components than protein, to assess how other pea components may impact the specific protein effects. Regardless of the differences in their structure, all supplement powders will be adjusted to provide the same amount of protein. The protein calculation will be done exclusively for each participant, based on their body weight (kg). They will consume up to 2 g of protein per body weight per day. Subjects will be asked to maintain their usual diet and their food intake will be accompanied during the intervention.
Participants' compliance will be followed by their food diaries and by clinical markers for protein intake and will collect blood, faeces and 24 h urine samples. A subset of the participants (max. 15 per arm) for visits 2 and 6 will be invited to participate in a postprandial challenge. After the fastening blood sample collection, they will receive the daily protein portion as breakfast. For those, 9 blood sample time points will be taken, being at 0, 15, 30, 60, 90, 120, 150, 180, and 480 min. After the 180 min measurement, they will receive a standard meal as lunch (e.g., pasta with tomato sauce).
研究设计
- 研究类型
- Interventional
- 分配方式
- Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- Triple (Participant, Investigator, Outcomes Assessor)
入排标准
- 年龄范围
- 18 Years 至 45 Years（Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Age 18-45 years
- •Body mass index (BMI) 18,5-30 kg/m2
- •Weight stable within the previous 3 months
- •Maintenance of the usual physical activity habits during the study
- •Intake of fibre between 15 and 25 g per day (evaluated by 3 food diaries or 24-hours recalls)
- •Omnivores
排除标准
- •Acute chronic disease, inflammatory or functional gastrointestinal diseases and any other disease or disorder that could affect the outcome of the study
- •Use of a medication that may interfere the study outcome
- •Eating disorder
- •High protein intake (more than 15% of energy or maximum 1,2 g of protein per kg of body weight per day; evaluated by 3 food diaries or 24-hours recalls)
- •Use of antibiotic medication during the last 3 months prior the first visit
- •Use of antibiotic medication very early in life (e.g., asthmatic children or ear inflammation)
- •Use of laxative or anti-diarrhoea medication within the past 3 months before the study
- •Regular consumption of probiotic or prebiotic product for the past 6 weeks before the study
- •Special diet that is considered to affect the study participation and/or study results, for example, high protein diets
- •More than 5 h of moderate-vigorous exercise per week
- •Pregnancy or breastfeeding
- •Intolerance to dietary supplements that will be used in the study
- •Abuse of alcohol or drugs
结局指标
主要结局
Changes in the faecal metabolites using targeted and untargeted metabolomics during the intervention
时间窗: 8 weeks
Difference in faecal metabolites depending on the protein supplementation amount and from colonic in vitro fermentation. Targeted and untargeted metabolomics measurements will be used (e.g., short-chain fatty acids and branched chain fatty acids analyses, and polar and non polar metabolomics). Faecal samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared throughout the whole study.
Changes in the plasma metabolites using targete and untargeted metabolomics during the intervention
时间窗: 4 weeks
Difference in plasma metabolites (blood obtained fastened state) depending on the protein supplementation amount. Targeted and untargeted metabolomics measurements will be used (e.g., lipids, and polar and non polar metabolomics). Plasma samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared throughout the whole study.
Colonic in vitro fermentation
时间窗: 4 weeks
Subjects will have their fresh faecal samples used in the colonic in vitro fermentation experiment to evaluate whether and how gut metabolites production can be predicted in vitro. The same proteins used in the dietary intervention study will be digested in vitro and used for the batch in vitro colonic fermentation. Supernatant will be used for metabolomics analysis (outcome 1) and selected fermentation time point pellets will be used for microbiota composition and functional analyses (outcome 3).
Profile/composition of the gut microbiota during the intervention
时间窗: 8 weeks
Difference in the gut microbiota profile/composition because of the protein supplementation and from colonic in vitro fermentation will be assessed by 16s/NGS. Alpha and Beta diversity will be measured and compared throughout the 4 weeks of intervention and baselines. In the case of NGS, the genome sequence will be annotated and the resulting set of genes encoding for metabolic enzymes will be extracted. Data from shotgun metagenomics and metabolomics will be integrated by constructing dependency networks, with a special attempt to infer causal relationships among different variables.
Routinely analysed markers for protein intake/compliance during the intervention - urine
时间窗: 4 weeks
Difference in 24 h urine samples markers due to the protein supplementation (e.g., for 24 h urine: urea nitrogen from 24 h urine, creatinine, uric acid)
Routinely analysed markers for protein intake/compliance during the intervention - blood
时间窗: 4 weeks
Difference in blood (serum) samples markers due to the protein supplementation (e.g., urea and creatinine)
Changes in the plasma metabolites using targeted and untargeted metabolomics during the dietary challenge
时间窗: 4 weeks
Difference in plasma metabolites depending on the protein taken as breaskfast. Targeted and untargeted metabolomics measurements will be used (e.g., polar and non polar metabolomics). Plasma samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared with those found in the fastening state.
次要结局
- Food intake using food diaries(8 weeks)
- Assessment of the bowel movement using the Bristol scale diary(8 weeks)
- Assessment of physical activity level(8 weeks)
- Assessment of the Gastrointestinal Symptom rating scale (GSRS) during the study(8 weeks)
- Assessment of height(4 weeks)
- Assessment of body weight during the study(4 weeks)
- Assessment of body composition(4 weeks)
- Fasting blood glucose level(4 weeks)
- Concentration of glucose - dietary challenge(4 weeks)
- Concentration of insulin - dietary challenge(4 weeks)
- Concentrations of faecal calprotectin(4 weeks)
- Fasting blood insulin level(4 weeks)
- Fasting blood C-reactive protein level(4 weeks)
- Fasting blood cholesterol (total, LDL, and HDL) level(4 weeks)
- Fasting blood triglycerides level(4 weeks)
- Changes in the faecal metabolites using targeted and untargeted metabolomics after the intervention(2 weeks)
- Breath test(4 weeks)
- Profile/composition of the gut microbiota after the intervention(2 weeks)