Dietary Pea Fiber Supplementation Improves Glycemia and Induces Changes in the Composition of Gut Microbiota, Serum Short Chain Fatty Acid Profile and Expression of Mucins in Glucose Intolerant Rats

Zohre Hashemi, Janelle Fouhse, Hyun Seun Im, Catherine B Chan, Benjamin P Willing, Zohre Hashemi, Janelle Fouhse, Hyun Seun Im, Catherine B Chan, Benjamin P Willing

Abstract

Several studies have demonstrated the beneficial impact of dried peas and their components on glucose tolerance; however, the role of gut microbiota as a potential mediator is not fully examined. In this study, we investigated the effect of dietary supplementation with raw and cooked pea seed coats (PSC) on glucose tolerance, microbial composition of the gut, select markers of intestinal barrier function, and short chain fatty acid profile in glucose intolerant rats. Male Sprague Dawley rats were fed high fat diet (HFD) for six weeks to induce glucose intolerance, followed by four weeks of feeding PSC-supplemented diets. Cooked PSC improved glucose tolerance by approximately 30% (p < 0.05), and raw and cooked PSC diets reduced insulin response by 53% and 56% respectively (p < 0.05 and p < 0.01), compared to HFD (containing cellulose as the source of dietary fiber). 16S rRNA gene sequencing on fecal samples showed a significant shift in the overall microbial composition of PSC groups when compared to HFD and low fat diet (LFD) controls. At the family level, PSC increased the abundance of Lachnospiraceae and Prevotellaceae (p < 0.001), and decreased Porphyromonadaceae (p < 0.01) compared with HFD. This was accompanied by increased mRNA expression of mucin genes Muc1, Muc2, and Muc4 in ileal epithelium (p < 0.05). Serum levels of acetate and propionate increased with raw PSC diet (p < 0.01). These results indicate that supplementation of HFD with PSC fractions can improve glycemia and may have a protective role against HFD-induced alterations in gut microbiota and mucus layer.

Keywords: glucose tolerance; gut microbiome; mucins; pea fiber; short chain fatty acids.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of 4 weeks of feeding a HFD supplemented with raw or cooked PSC on oral glucose tolerance test (OGTT) and the corresponding insulin response. (a) Concentrations of blood glucose measured basally and following oral administration of 1 g/kg glucose; (b) IAUC calculated for glucose levels during OGTT; (c) Plasma insulin levels during OGTT; (d) incremental area under the curve (IAUC) for insulin during OGTT. The data represent mean ± SEM, n = 4–14. Significant differences seen at different time points of OGTT are explained in the text, and differences between IAUCs are shown here. * p < 0.05, ** p < 0.01 and *** p < 0.001 show significant difference compared with HFD. Reproduced from Hashemi et al. [6] with permission.
Figure 2
Figure 2
Effect of feeding PSC-supplemented diets on fecal microbial composition. (a) Fecal bacterial communities clustered using PCoA analysis of weighted UniFrac distances, analyzed by ANOSIM. The percentage of variation explained by each coordinate is shown in parentheses. HFD and LFD clustered together (p > 0.05) and cooked (CP) and raw (RP) clustered together (p > 0.05); however, HFD and LFD clustered separately from CP and RP (p < 0.05); (b) Inverse Simpson diversity index as a measure of diversity within each sample. Inverse Simpson diversity indices differed significantly among the groups (p < 0.001). Bars are means ± SEM, n = 6–8, ** p < 0.01, * p < 0.05.
Figure 3
Figure 3
mRNA expression of TLRs and tight Junction proteins. Mean relative mRNA expression (FC, fold change) of (a) TLR2; (b) TLR4; (c) occludin and (d) ZO-1 in ileum of the rats normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. Data are means ± SEM, n = 5–8. No significant differences were found between groups.
Figure 4
Figure 4
mRNA expression of mucins. Relative mRNA expression (FC, fold change) of (a) Muc1; (b) Muc2; (c) Muc3 and (d) Muc4 in the ileum. Gene expression data was normalized to GAPDH as the house-keeping gene and presented as means ± SEM, n = 6–8. There was a significant effect of treatment for relative expression of Muc1, Muc2, and Muc4 (p < 0.05). CP group showed increased expression of Muc2 and Muc4 genes when compared to HFD group (* p < 0.05).
Figure 5
Figure 5
SCFA concentration (μmol/g) in (a) serum and (b) feces of rats following four weeks of PSC-supplemented or control diets. Data are presented as means ± SEM, n = 6–8. Serum acetate and propionate concentrations were significantly different between the groups. Asterisks show significant difference compared to HFD (** p < 0.01, * p < 0.05). Fecal SCFAs concentrations were not statistically different between groups.

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