ABSTRACT
The intracellular sensor NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome controls caspase-1 activity and the maturation and release of the cytokines interleukin (IL)-1ß and IL-18. The NLRP3 inflammasome has attracted the attention of the pharmaceutical industry because it promotes the pathogenesis of many diseases, making it a promising target for drug development. Litsea cubeba (Lour.) is a plant traditionally used as a seasoning in Taiwan and in other Asian countries. In this study, we investigated the inhibitory activity of the leaves of L. cubeba against the NLRP3 inflammasome. We found that the ethanol extract of L. cubeba leaves (MLE) inhibited the NLRP3 inflammasome in macrophages by reducing caspase-1 activation and IL-1ß secretion. MLE reduced pyroptosis in macrophages and inhibited the release of NLRP3 and apoptosis-associated speck-like protein containing a CARD (ASC). In a mechanistic study, MLE reduced mitochondrial reactive oxygen species (ROS) production and preserved mitochondrial integrity, which led to reduced mitochondrial DNA release into the cytosol. MLE did not reduce the expression levels of NLRP3, IL-1ß precursor or TNF-α in lipopolysaccharide (LPS)-activated macrophages. These results indicated that MLE inhibited the NLRP3 inflammasome by suppressing the activation signals of the NLRP3 inflammasome but not by reducing the priming signal induced by LPS. In addition, oral administration of MLE (20-80 mg/kg) ameliorated dextran sulfate sodium (DSS)-induced colitis in a mouse model. Notably, mice that received MLE (1 and 2 g/kg) daily for 7 days did not exhibit visible side effects. Gas chromatography-mass spectrometry (GC-MS) analysis found that α-Terpinyl acetate (27.2%) and 1,8-Cineole (17.7%) were the major compounds in MLE. These results indicated that L. cubeba leaves have the potential to be a nutraceutical for preventing and improving NLRP3 inflammasome-related diseases.
ABSTRACT
Prophylactic use of antibiotics in-feed has been effective in decreasing the incidence of diarrhea in weaning piglets. However, the overuse of antibiotics as prophylactic or therapeutic agents in animal feed leads to the evolution of drug-resistant bacteria and antibiotic residues in pigs. This study investigated the effects of Bacillus licheniformis-fermented products on diarrhea incidence and the fecal microbial community in weaning piglets. A total of 120 crossbred piglets with an average initial body weight of 9.87 ± 1.43 kg were randomly allotted to four dietary treatments consisting of three replicate stalls with 10 piglets in each. The dietary treatments comprised a basal diet as control, control plus 1 g/kg or 4.5 g/kg of B. licheniformis-fermented products, and control plus 30 mg/kg antibiotics (bacitracin methylene disalicylate). Results showed that 4.5 g/kg of B. licheniformis-fermented product supplementation significantly reduced diarrhea incidence in weaning piglets. Principal coordinate analysis and a heatmap of species abundance indicated distinct clusters between the groups treated with antibiotics and B. licheniformis-fermented products. The bacterial richness and evenness in the feces decreased in weaning piglets fed 1 g/kg of B. licheniformis-fermented products and antibiotics. The abundance of the genera [Ruminococcus] gauvreauii group, Ruminococcaceae UCG-005, and Ruminococcaceae UCG-008 in the feces decreased in weaning piglets fed B. licheniformis-fermented products or antibiotics. The average abundance of the genus Prevotella 9 in the feces was positively correlated with the concentration of B. licheniformis-fermented products and negatively correlated with the diarrhea incidence in weaning piglets. Furthermore, the average abundance of the genus Prevotella 9 in the feces was positively correlated with the growth performance of weaning piglets. These results demonstrate that B. licheniformis-fermented products can improve diarrhea incidence and fecal microflora composition in weaning piglets.
