The objective of this experiment was to investigate the influence of weaning stress and an antioxidant blend on gut health and free radical metabolism in postweaning pigs. A total of 96 pigs from 12 litters were randomly divided by litter to 3 groups with 4 litters each. The control group and the weaning group were fed the basal diet, and the antioxidant group was fed the basal diet supplemented with an antioxidant blend. The control group was suckling normally during the experimental period and the other 2 treatments were weaned at 21 d of age. Morphology in different parts of the intestines was used as a measure of intestinal barrier function. Activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and the concentrations of malondialdehyde (MDA), NO, H₂O₂, and O₂ were measured in serum. Activities of the digestive enzymes, including sucrase, maltase, amylase, lipase, and pepsin, were measured at 24 d of age for all treatments. Gene expressions of free radicals, digestive enzymes, or antioxidant enzymes were selected for quantitative reverse transcription-PCR analyses. Results showed that weaning resulted in reductions (P < 0.05) in the villus height and width, and activity of digestive enzymes. Activity of SOD decreased (P < 0.05) and the concentrations of MDA, NO, and H₂O₂ increased (P < 0.05) after weaning. The expression results indicated that the genes related to the antioxidant enzymes and digestive enzymes were down regulated (P < 0.05) after weaning. Tumor protein 53, which regulates reactive oxygen-species generation, tended to increase (P < 0.10) in the weaning group. The concentration of PPARγ coactivator-1α (PGC-1α), which plays an important protective role against oxidative stress by regulating the expression of mitochondrial antioxidants, was reduced (P < 0.05) in weaning pigs and increased (P < 0.01) in antioxidant pigs compared with the control pigs. Results indicated that intestinal dysfunction occurred after weaning and there was an inhibition of the antioxidant system. The antioxidant blend has the potential to prevent free radical-induced damage and suppress oxidative stress by modulating the expressions of tumor protein 53 and PGC-1α genes.

Lactobacillus spp., as probiotics, have shown efficacy in alleviating nonalcoholic fatty liver disease (NAFLD). Here, we screened a new probiotic strain, Lactobacillus salivarius SNK-6 (L. salivarius SNK-6), which was isolated from the ileum of healthy Xinyang black-feather laying hens in China. We investigated the beneficial activity of L. salivarius SNK-6 in a NAFLD model in laying hens and found that L. salivarius SNK-6 inhibited liver fat deposition and decreased serum triglyceride levels and activity of aspartate transaminase and alanine transaminase. MBOAT2 (membrane-bound O-acyltransferase domain containing 2) was directly targeted by miR-130a-5p, which was downregulated in the liver of NAFLD laying hens but reversed after L. salivarius SNK-6 treatment. Downregulation of MBOAT2, L. salivarius SNK-6 supplementation in vivo, and L. salivarius SNK-6 cell culture treatment in vitro suppressed the mRNA expression of genes involved in the PPAR/SREBP pathway. In addition, 250 metabolites were identified in the supernatants of L. salivarius SNK-6 culture media, and most of them participated in metabolic pathways, including amino acid, carbohydrate, and lipid metabolism. Targeted metabolomic analysis revealed that acetate, butyrate, and propionate were the most abundant short-chain fatty acids, while cholic acid, ursodeoxycholic acid, chenodeoxycholic acid, and tauroursodeoxycholic acid were the four most-enriched bile acids among L. salivarius SNK-6 metabolites. This may have contributed to the reparative effect of L. salivarius SNK-6 in the NAFLD chicken model. Our study suggested that L. salivarius SNK-6 alleviated liver damage partly via the miR-130a-5p/MBOAT2 signaling pathway.

Many studies have shown that Orthosiphon stamineus extract (OE) has antioxidant activity, and we previously reported that OE protects the intestine against injury from a high-fat diet. However, the molecular mechanism underlying this protective effect of OE was unclear. Here, OE was separated according to polarity and molecular weight, and the antioxidant activity of each component was compared. The components with the highest antioxidant activity were analyzed by HPLC, which confirmed that rosmarinic acid (RA) was the main effective constituent in OE. OE and RA were then tested in a mouse high-fat diet-induced intestinal injury model. The antioxidant indices and morphological characteristics of the mouse jejunum were measured, and activation of the nuclear factor E2-related factor 2 (Nrf2) pathway and apoptosis of jejunal epithelial cells were analyzed. Of all the constituents in OE, RA contributed the most. Both RA and OE activated the Nrf2 pathway and increased downstream antioxidant enzyme activity. RA and OE protected the mouse intestine against high-fat diet-induced oxidative stress by preventing intestinal epithelial cell apoptosis via both extracellular and intracellular pathways. Thus, RA, the main effective constituent in OE, inhibits intestinal epithelial apoptosis by regulating the Nrf2 pathway in mice.