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. 2022 Oct 14;14(20):4301.
doi: 10.3390/nu14204301.

Milk Formula Enriched with Sodium Butyrate Influences Small Intestine Contractility in Neonatal Pigs

Affiliations

Milk Formula Enriched with Sodium Butyrate Influences Small Intestine Contractility in Neonatal Pigs

Monika Słupecka-Ziemilska et al. Nutrients. .

Abstract

Butyrate, a by-product of gut bacteria fermentation as well as the digestion of fat in mother's milk, exerts a wide spectrum of beneficial effects in the gastrointestinal tissues. The present study aimed to determine the effects of sodium butyrate on small intestine contractility in neonatal piglets. Piglets were fed milk formula alone (group C) or milk formula supplemented with sodium butyrate (group B). After a 7-day treatment period, isometric recordings of whole-thickness segments of the duodenum and middle jejunum were obtained by electric field stimulation under the influence of increasing doses of Ach (acetylocholine) in the presence of TTX (tetrodotoxin) and atropine. Moreover, structural properties of the intestinal wall were assessed, together with the expression of cholinergic and muscarinic receptors (M1 and M2). In both intestinal segments (duodenum and middle jejunum), EFS (electric field stimulation) impulses resulted in increased contractility and amplitude of contractions in group B compared to group C. Additionally, exposure to dietary butyrate led to a significant increase in tunica muscularis thickness in the duodenum, while mitotic and apoptotic indices were increased in the middle jejunum. The expression of M1 and M2 receptors in the middle jejunum was significantly higher after butyrate treatment. The results indicate increased cholinergic signaling and small intestinal growth and renewal in response to feeding with milk formula enriched with sodium butyrate in neonatal piglets.

Keywords: cholinergic signaling; intestinal contractility; neonates; pigs; sodium butyrate.

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Conflict of interest statement

The authors disclose no conflict of interest.

Figures

Figure 1
Figure 1
The thickness of the muscularis mucosa muscle layer (μm) in the duodenum and middle jejunum of control (C) and sodium-butyrate-treated piglets (B). Values are given as means ± SD; * indicates statistical differences between groups. In the duodenum, sodium butyrate led to a significant increase in the thickness of the muscle layer (**** p < 0.0001). On the contrary, a significant reduction in the muscularis thickness was observed in the middle jejunum after sodium butyrate administration (** p = 0.0023).
Figure 2
Figure 2
Amplitude of contractions (mm) of isolated duodenal and middle jejunum fragments in control (C) and sodium-butyrate-treated piglets (B). Values are given as means ± SD; * indicates statistical differences between groups when * p < 0.05, *** p < 0.001.
Figure 3
Figure 3
Representative microscopic images of the middle jejunum mucosa in 10-day-old control (C) and sodium-butyrate-treated piglets (B) (at a dose of 0.3%), with apoptotic cells (red) visualized using a TUNEL assay. Intestinal villi (A) and crypts (B) in piglets from the control group (group C). Intestinal villi (C) and crypts (D) in piglets from the sodium butyrate treated group (group B). The tissue structure was visualized using Nomarski contrast in a confocal microscope (LSM Pascal, Zeiss, Germany, magnification: 400×). Ki67-positive epithelial cells (3,3′-diaminobenzidine staining, brown) in crypts counterstained with hematoxylin in blue in animals from the control group (group C) (E) and from the sodium-butyrate-treated group (group B) (F). Light microscope (Axioskop 40, Zeiss, Germany), magnification: 400×.
Figure 4
Figure 4
Quantitative results of optical density and representative Western blots of M1, M2, and M3 receptor expression in the lysates from the mucosa of the middle jejunum segments in newborn pigs. Piglets fed milk formula alone (group C) or supplemented with sodium butyrate at a dose of 0.3% (group B). Values are given as means ± SEM; * indicates statistical differences between groups when ** p < 0.01, **** p < 0.0001.

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