Effects of indigestible carbohydrates in barley on glucose metabolism, appetite and voluntary food intake over 16 h in healthy adults

The metabolic syndrome (MetS) represents a cluster of risk factors identifying subjects at high risk of developing type 2 diabetes (T2D) and cardio-vascular disease [1]. The prevalence of obesity and T2D is increasing globally and the World Health Organization estimates in 2012 the number of people suffering from T2D worldwide to be 347 million, and that the number of diabetes deaths will increase by two thirds between year 2008 and 2030 [2]. The need for preventive strategies is thus urgent.

Observational studies have shown that higher intake of whole grain (WG) is associated with lower body mass index (BMI) [3‐5], improved insulin sensitivity [6], reduced risk of T2D [7] and of premature total and cause-specific death [8]. Several mechanisms have been discussed as mediators of favorable metabolic outcomes following WG intake, e.g. low GI features, presence of dietary fibre (DF) per se and/or the presence of DF associated bioactive components [9‐12]. It has been hypothesized that dietary factors may affect composition and metabolism of the gut microflora [13, 14], and a close connection between the microbial composition and inflammatory status has been observed [15, 16]. Consequently, metabolic derangement and also obesity has been described as an endotoxemic inflammatory condition triggered by e.g. high fat feeding [17, 18]. Oligofructose has been shown to increase bifidobacteria in obese mice [19], which was accompanied by improved glucose regulation and a reduced inflammatory tonus [20, 21], indicating a prebiotic mechanism. Less is known about possible colonic mediated effects of DF present in WG diets. A relation between WG intake and improved inflammatory status was reported in a cross-sectional study in healthy subjects [22]. In addition, certain WG products, rich in cereal DF and resistant starch (RS) facilitated blood glucose regulation and improved inflammatory tonus in humans in the perspective from a late evening meal to a standardized breakfast, i.e. in the time frame 10 to 12 h after meal [23]. The metabolic benefits has been proposed to be associated with colonic fermentation of indigestible carbohydrates (DF and RS) [24‐26], and an associated increase in systemic levels of glucagon-like peptide-1 (GLP-1) [23]. GLP-1 is increasingly being assigned both anti-diabetic and anti-obesogenic features [27‐29], which makes it relevant to perform further studies regarding the possible relation between food derived stimulation of this incretin and effects on metabolic parameters.

The objective of this work was to evaluate the effect of intrinsic indigestible carbohydrates in boiled barley kernels (BK) consumed in the evening on glycaemia, appetite sensations, appetite regulatory hormones and voluntary food intake, at fasting and in the post-prandial phase following two consecutive meals (breakfast and lunch). Variables determined in blood were glucose, insulin, GLP-1, glucose-dependent insulinotropic polypeptide (GIP), ghrelin, free fatty acids (FFA), adiponectin and interleukin (IL)-6. In addition breath hydrogen (H₂) excretion was determined as a marker of colonic fermentation. The breakfast and lunch meals were provided ad libitum, allowing for evaluation of over-night effects on metabolism and appetite regulation in a realistic eating situation. For this purpose, carbohydrate-equivalent meals, consisting of boiled BK or white wheat bread (WWB, reference meal), were provided as evening meals to healthy subjects, using a randomized cross-over design.

The purpose of the present study was to examine the metabolic effects of intrinsic fermentable carbohydrates in boiled barley kernels in an over-night perspective. In comparison to previous studies [23, 35], the experimental design is novel in that the experimental sampling period is significantly extended, covering a period ranging from 10.5-16 h after ingestion of test and reference evening meal. Further the present study aimed to investigate the influence of evening meals on metabolic test variables, appetite, and voluntary energy intake following ad libitum breakfast- and lunch meals; that is in a more realistic eating situation than studies with standardized meal size.

Previous studies have revealed that dietary supplementation of inulin and oligofructan during two weeks [28], or consumption of other specific indigestible carbohydrates, e.g. present in BK, as a single late evening meal [23, 36], decrease the glucose response and increase gut microbial activity (as determined by breath H₂ and/or plasma short chain fatty acids (SCFA)) at the following breakfast meal in healthy humans. In accordance with previous findings, the present study revealed decreased glucose response after breakfast following BK as an evening meal compared to WWB, and increased gut microbial fermentation, as indicated by elevated breath H₂ concentrations. As a novel finding, the BK evening meal decreased glucose iAUC during the course of the entire experimental day (0–330 min), and increased GP² after breakfast, all indicative of smoother postprandial glucose excursion. In this context it must be noted that in contrast to the standardized meal size previously used, glucose measurements in the present study were conducted after ad libitum food intake (standardized in quality). At breakfast there was only a minor (4%, non-significant) decrease in energy intake after BK. It is therefore highly unlikely that the differences seen in metabolic variables after breakfast could be explained by differences in energy intake at breakfast. However, the glucose responses at lunch following the BK evening meal may, at least to a minor degree, have been affected by the reduced caloric intake at lunch.

It has been hypothesized that modulation of gut microbiota by oligofructans, may interfere beneficially with host metabolism. Thus, dietary supplementation with oligofructose for 14 weeks decreased glucose AUC and voluntary energy intake, as well as reduced inflammatory tonus in mice models [21]. Studies in rodents have further shown that oligofructose feeding during 4–5 weeks may promote epithelial L-cell differentiation in the gut, contributing to higher GLP-1 production [20, 37]. It has been demonstrated that SCFA produced by bacterial fermentation may trigger signaling cascades through acting on SCFA receptors on L-cells (in vitro model), resulting in increased release of gut peptides such as GLP-1 and Peptide YY (PYY) [38]. Fructan supplementation during two weeks was also reported to reduce glucose response, reduce feeling of hunger and increase GLP-1 concentration in response to an ad libitum meal in humans [28]. Less is known about the gut mediated effects of intrinsic indigestible carbohydrates in food, e.g. boiled barley kernels, on glycaemia, gut fermentation, and appetite regulation, and to the authors’ knowledge, data from human studies are scarce. Nilsson et al. (2008) found inverse correlations between glucose iAUC and breath H_2, supporting a link between colonic fermentation and glucose regulation [23]. A particularly interesting observation in the present study is a 34% increase in plasma concentration of GLP-1 in the morning (0–330 min) after the BK evening meal, compared to the evening meal with WWB. In addition, in the time period 60–120 min after breakfast also GIP concentrations (AUC) were increased after BK, which is in accordance with previous findings in our research group [23]. Both GLP-1 and GIP have implications in glucose homeostasis, and in addition GLP-1 is considered as a modulator of appetite regulation. Both glucose intolerance and obesity seems to be related to decreased levels of GLP-1, independently of one another [27]. In the present study, the BK evening meal decreased the feeling of hunger during the whole experimental day, as compared to the evening WWB, and reduced voluntary energy intake (−12%) at lunch. Previously it has been found that intravenous infusions of GLP-1 (50 pmol/kg⋅h) during 4 h reduced voluntary energy intake by 12% at a subsequent lunch meal, as compared to saline in young healthy men [39]. The results in the present study are thus in accordance with previous studies showing that GLP-1 reduce food intake and increase satiety in both lean and obese subjects [40]. Although not significant, there was a trend (P = 0.07) to reduced fasting levels of ghrelin after the BK evening meal by 16% and there was a, non-significant, 10% reduction of ghrelin in the late post-prandial phase prior to lunch (AUC 120–210 min) as compared to after WWB. Intravenous administration during 4 h of the orexigenic hormone ghrelin, demonstrates increased food intake in healthy subjects at a subsequent meal [41]. A connection has been proposed between colonic fermentation and reduced concentrations of serum ghrelin in healthy subjects, 6 h after ingestion of inulin, possible mediated through the formation of SCFA [42]. It can be suggested that the effects on gastro-intestinal hormones seen in the present study after the BK evening meal, stem from colonic fermentation of indigestible DF in the BK. GLP-1 and GIP, are known to exert insulinotropic effects [43] and have been suggested to account for up to 70% of meal induced insulin release in humans [44]. Noticeably, the lowered glycaemia and increased incretin concentrations observed during the experimental day after BK evening meal were not accompanied by increased insulin response. However, GLP-1 has previously been reported to improve insulin sensitivity in T2D [45], a phenomenon that might have contributed to the facilitated glucose regulation during the experimental day in the present study in healthy subjects. Another factor that might have contributed to an improved insulin sensitivity after the BK evening meal is reduced f-FFA concentrations compared to the evening WWB reference meal [46]. Belfort et al. (2005) showed that even a modest increase in plasma FFA, well within the physiological range, causes a dose-dependent inhibition of insulin-stimulated glucose disposal and insulin signaling in healthy lean subjects [47].

In the present study, f-IL-6 concentrations tended to be decreased (P = 0.06) after the BK evening meals, which is in accordance with previous studies [23, 36]. Low-grade chronic systemic inflammation is associated with obesity and insulin resistance [48]. Anti-inflammatory properties of specific indigestible carbohydrates, as present in e.g. BK, might constitute a promising approach aiming at dietary prevention and/or treatment of obesity and the metabolic syndrome. In a cross-sectional study by Yannakoulia et al. (2008), consumption of WG cereals and low-fat dairy products were positively associated with adiponectin concentrations among healthy women [49]. The authors suggested that adiponectin may be a mediator of the decreased risk of T2D associated with higher WG intake. Adiponectin has been shown to possess not only insulin sensitizing properties but also to have anti-inflammatory effects [50]. Decreased concentrations of adiponectin are observed in obesity and T2D [51, 52]. In the present study, we show that BK evening meal resulted in maintained adiponectin concentrations in the post-prandial period after breakfast as compared to the decrease observed after WWB evening meal. The metabolic relevance of this finding remains to be elucidated. However, adiponectin concentrations in subjects with metabolic disorders show a more pronounced decrease in the post-prandial phase compared to healthy subjects [53, 54].

The results indicate that indigestible carbohydrates, as present in BK, have the potential to facilitate glucose regulation in healthy subjects in a time period of 10.5-16 h, decrease inflammatory markers, decrease FFA, decrease hunger sensations and reduce energy intake at a subsequent lunch. Interestingly, the BK evening meal resulted in an increased release of GLP-1 during the whole experimental period. The effects are suggested to be mediated through gut microbial fermentation, proposing a role of gut microbiota in modulating host metabolism in humans. Colonic fermentation of specific indigestible carbohydrates may provide one possible mechanism by which WG have proven beneficial in prevention of obesity and T2D. Taken together, the beneficial effects of BK are supportive for a prebiotic potential of intrinsic indigestible carbohydrates in barley kernel based products.

ACN, EMÖ, and IMEB designed the research; EVJ, ACN, EMÖ, and IMEB supervised and/or conducted the research; EVJ analyzed data and performed statistical analysis; EVJ, ACN, EMÖ, and IMEB wrote the paper; and ACN, EMÖ and IMEB had primary responsibility for the final content. All authors have read and approved the final manuscript.