Abstract

β-Conglycinin (β-CG), an anti-nutritional factor, is a major allergen in soybeans to induce intestinal dysfunction and diarrhea in neonatal animals, including piglets and human infants. This study with a piglet model determined the effects of N-acetylcysteine (NAC) on intestinal function and autophagy in response to β-CG challenge. Twenty-four 12-day-old piglets (3.44 ± 0.28 kg), which had been weaned at 7 days of age and adapted for 5 days after weaning, were randomly allocated to the control, β-CG, and β-CG + NAC groups. Piglets in the control group were fed a liquid diet containing 10% casein, whereas those in the β-CG and β-CG + NAC groups were fed the basal liquid diets containing 9.5% casein and 0.5% β-CG for 2 days. Thereafter, pigs in the β-CG + NAC group were orally administrated with 50 mg (kg BW)−1 NAC for 3 days, while pigs in the other two groups were orally administrated with the same volume of sterile saline. NAC numerically reduced diarrhea incidence (− 46.2%) and the concentrations of hydrogen peroxide and malondialdehyde, but increased claudin-1 and intestinal fatty-acid binding protein (iFABP) protein abundances and activities of catalase and glutathione peroxidase in the jejunum of β-CG-challenged piglets. Although β-CG challenge decreased the villus height, villus height/crypt depth ratio, and mRNA levels of claudin-1 and occludin, no significant differences were observed in these indices between the control and β-CG + NAC groups, suggesting the positive effects of NAC supplementation on intestinal mucosal barrier function. Moreover, NAC increased the concentrations of citrulline and D-xylose in the plasma, as well as the expression of genes for aquaporin (AQP) 3, AQP4, peptide transporter 1 (PepT1), sodium/glucose co-transporter-1 (SGLT-1), potassium inwardly-rectifying channel, subfamily J, member 13 (KCNJ13), and solute carrier family 1 member 1 (SLC1A1) in the jejunum, demonstrating that NAC augmented intestinal metabolic activity and absorptive function. Remarkably, NAC decreased Atg5 protein abundance and the LC3II/LC3I ratio (an indicator of autophagy) in the jejunum of β-CG-challenged piglets. Taken together, NAC supplementation improved intestinal function and attenuated intestinal autophagy in β-CG-challenged piglets.

Highlights

  • Soybean is a high-quality protein source, and possesses good nutritional, processing, and functional p­ roperties[1]

  • Given the oxidative stress caused by β-CG a­ dministration[7], we proposed that reactive oxygen species (ROS) play a critical role in elevating autophagy activity in enterocytes

  • Using a porcine or chicken model of intestinal dysfunction induced by lipopolysaccharide or heat stress, we found that dietary NAC supplementation could improve intestinal function and animal ­health[16,17]

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Summary

Results

Β-CG challenge increased (P < 0.05) the incidence of diarrhea in piglets, compared with the control group. NAC supplementation attenuated the reductions in the mRNA levels for jejunal claudin-1 and occludin of β-CG-challenged piglets based on the finding that there were no significant differences in the expression of these two genes between the control and β-CG + NAC groups. 7 and 8, β-CG challenge increased (P < 0.05) the abundance of beclin-1 and HSP70 proteins, and the LC3II/LC3I ratio, while decreasing (P < 0.05) the abundance of AQP3, iFABP, claudin-1, and occludin proteins in the jejunum of piglets. No significant differences were observed in the protein abundances of AQP3 or LC3II/LC3I ratio in the jejunum between the control and β-CG + NAC groups of piglets No significant differences were observed in the protein abundances of AQP3 or LC3II/LC3I ratio in the jejunum between the control and β-CG + NAC groups of piglets (Figs. 7 and 8), indicating that NAC attenuated the decreases in these two variables in β-CG-challenged piglets

Discussion
Conclusions
Materials and methods
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