Hesperidin Effects on Gut Microbiota and Gut-Associated Lymphoid Tissue in Healthy Rats

Sheila Estruel-Amades, Malén Massot-Cladera, Francisco J Pérez-Cano, Àngels Franch, Margarida Castell, Mariona Camps-Bossacoma, Sheila Estruel-Amades, Malén Massot-Cladera, Francisco J Pérez-Cano, Àngels Franch, Margarida Castell, Mariona Camps-Bossacoma

Abstract

Hesperidin, found in citrus fruits, has shown a wide range of biological properties. Nonetheless, a more in-depth investigation is required on the effects on the immune system, and in particular, on the gut-associated lymphoid tissue, together with its relationship with the gut microbiota. Therefore, we aimed to establish the influence of oral hesperidin administration on the intestinal lymphoid tissue and on the gut microbiota composition in healthy animals. Lewis rats were orally administrated 100 or 200 mg/kg hesperidin three times per week for four weeks. Microbiota composition and IgA-coated bacteria were determined in caecal content. Mesenteric lymph node lymphocyte (MLNL) composition and functionality were assessed. IgA, cytokines, and gene expression in the small intestine were quantified. Hesperidin administration resulted in a higher number of bacteria and IgA-coated bacteria, with changes in microbiota composition such as higher Lactobacillus proportion. Hesperidin was also able to increase the small intestine IgA content. These changes in the small intestine were accompanied by a decrease in interferon-γ and monocyte chemotactic protein-1 concentration. In addition, hesperidin increased the relative proportion of TCRαβ+ lymphocytes in MLNL. These results show the immunomodulatory actions of hesperidin on the gut-associated lymphoid tissue and reinforce its role as a prebiotic.

Keywords: flavanone; flavonoids; immunoglobulin A; intestinal immunity; polyphenol; prebiotic.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of hesperidin administration on total polyphenol content in urine. Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group, δp < 0.05 versus H100 group (one-way ANOVA).
Figure 2
Figure 2
Effect of hesperidin administration on bacterial groups determined by FISH-FCM in caecal homogenates. (a) Counts of total bacteria; (b) counts of IgA-coated bacteria; (c) proportion of IgA-coated bacteria; proportions of (d) Streptococcus spp.; (e) Clostridium coccoides/Eubacterium rectale; (f) Lactobacillus/Enterococcus; (g) Sstaphylococcus spp.; (h) Clostridium histolyticum/Clostridium perfringens; (i) Bacteroides/Prevotella; (j) Bifidobacterium; and (k) Escherichia coli. Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group (one-way ANOVA).
Figure 3
Figure 3
Effect of hesperidin administration on the proportion of mesenteric lymph node lymphocytes. (a) TCRαβ+, TCRγδ+ and CD45RA+ lymphocytes; (b) TCRαβ+/CD45RA+ ratio; (c) Th (TCRαβ+CD4+) and Tc (TCRαβ+CD8+) lymphocytes; and (d) Th/Tc ratio. Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group (one-way ANOVA).
Figure 4
Figure 4
Effect of hesperidin administration on (a) the mesenteric lymph node lymphocyte (MLNL) proliferative activity; and (b–f) cytokine release by MLNL after anti-CD3/CD28 stimulation. Data are expressed as mean ± standard error (n = 5–6).
Figure 5
Figure 5
Effect of hesperidin administration on the cytokine concentration in the small intestine washes (a–e). Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group (Mann–Whitney U test).
Figure 6
Figure 6
Effect of hesperidin administration on (a) IgA concentration in faeces collected throughout the study; (b) caecum content; (c) small intestine wash; and on (d) IgM concentration in the small intestine wash. Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group (one-way ANOVA).
Figure 7
Figure 7
Effect of hesperidin administration throughout the study on serum (a) IgG; (b) IgA; and (c) IgM. Data are expressed as mean ± standard error (n = 5–6). Statistical difference: * p < 0.05 versus REF group, δp < 0.05 versus H100 group (Mann–Whitney U test).

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