Effect of feeding regimen on circadian activity rhythms of food anticipatory by ghrelin hormone in a pig model.

Increasingly diverse meal patterns affect the internal body clock. Ghrelin secretion is closely associated with the anticipation of a regularly scheduled mealtime, leading ghrelin to be a putative candidate for food-related entraining signals that drive activity rhythms. Here, growing pigs with different meal frequencies were used to construct an irregular eating pattern model. We found that irregular eating patterns changed central ghrelin levels of pigs, affected the circadian entrainment and circadian rhythm pathways in hypothalamus tissue, and altered the daily behavior and food anticipatory activity (FAA). To determine whether ghrelin exerts an effect, growing pigs were intravenously injected with ghrelin antagonist [D-Lys3]-GHRP-6 for 7 days. We showed here that [D-Lys3]-GHRP-6 administration decreased locomotor activity of growing pigs in the 4-h window preceding onset of food availability. In addition, we also confirmed that the direct role of ghrelin in molecular mechanism of regulating clock genes expression via calcium mobilization through intracellular PKC/PLC and AC/PKA pathways in vitro. Collectively, irregular eating patterns affect the central circadian system by ghrelin, supporting ghrelin as a temporal messenger of food-entrainment in hypothalamic circadian functions.

The development of intestinal lactic acid bacteria in piglets as determined by high-throughput sequencing.

The present study investigated the development of intestinal lactic acid bacteria (LAB) in piglets by high-throughput sequencing. Fresh feces from newborn piglets were collected at the ages of 1, 2, 3, 7, 14, 21, 24, 35 and 42 days. Sequencing of DNA amplicons of bacteria in feces amplified with LAB-specific primers showed that Lactobacillus was the predominant genus with an abundance higher than 70%, but tended to decrease after weaning (p = 0.059). The abundance of Streptococcus was the second predominant genus on day 1, then decreased significantly (p < 0.05). The relative abundance of Facklamia, Bacillus and Aerosphaera increased significantly after weaning (p < 0.05). At the species level, Lactobacillus reuteri, Uncultured Lactobacillus sp. and Lactobacillus mucosae dominated on day 3. Lactobacillus reuteri was the predominant bacteria at 7, 21, 24 and 35 days of age, and Lactobacillus sp. MWBPC 1-3-1 dominated on day 42. The relative abundance of Lactobacillus vaginalis, Lactobacillus johnsonii and Lactobacillus gasseri on day 24 was significantly higher than other ages (p < 0.05). These results indicate that the structure and predominant species of LAB changed dramatically from birth to weaning, which is of great significance to explore the LAB for the health of piglets.

Time-restricted feeding affects transcriptomic profiling of hypothalamus in pigs through regulating aromatic amino acids metabolism.

BACKGROUND: Time-restricted feeding (TRF) is an effective means that can efficiently regulate the metabolism and health of animals and humans. However, the effect of TRF on hypothalamic function remains unclear. RESULTS: Results showed that TRF significantly increased the activities of digestive enzymes lipase, maltase in the duodenum and lipase, trypsin in the pancreas whereas significantly decreased serum gastrointestinal hormones gastrin, glucagon-like peptide-1, cholecystokinin, peptide YY, and ghrelin. Metabolites related to amino acid metabolism, including citrulline, kynurenine, N-acetylleucine, l-tryptophan, and l-tyrosine, significantly increased in the TRF group. Differential metabolites were mainly enriched in phenylalanine, tyrosine, and tryptophan biosynthesis and tryptophan metabolism. Transcriptomic analysis of hypothalamus showed that a total of 462 differentially expressed genes (DEGs) were significantly changed by TRF. In particular, DEGs such as DDC, TH, GOT2, and DBH involved in aromatic amino acid metabolism pathways were significantly downregulated, whereas the expression of CYP1B1 was significantly upregulated. Moreover, DEGs (PDYN and PPP3CA) involved in amphetamine addiction and cocaine addiction were also downregulated in the TRF group. CONCLUSION: Taken together, these results suggested that TRF improved the digestion and absorption of nutrients and thus increased the accessibilities of aromatic amino acids. The increasing of circulating aromatic amino acids might mediate the regulatory neuroendocrine effects of TRF regimes on the hypothalamus functions, especially on drug addictions. This study reveals a possible mechanism underlying the effects of regulating feeding patterns on the function of the hypothalamus by altering aromatic amino acids metabolism. © 2022 Society of Chemical Industry.

Inhibition of ghrelin activity by the receptor antagonist [D-Lys3]-GHRP-6 enhances hepatic fatty acid oxidation and gluconeogenesis in a growing pig model.

Despite its central role in regulating energy intake and metabolism, ghrelin is little understood when it comes to its effects on hepatic lipid and glucose metabolism. Growing pigs were intravenously injected with ghrelin receptor antagonist [D-Lys3]-GHRP-6 (DLys; 6 mg/kg body weight) for seven days to determine whether ghrelin plays a role in glucose and lipid metabolism. DLys treatment significantly reduced body weight gain and adipose histopathology found that DLys treatment dramatically reduced adipocyte size. DLys treatment significantly increased serum NEFA and insulin levels, hepatic glucose level and HOMA-IR, and significantly decreased serum TBA level of growing pigs after fasting. Moreover, DLys treatment changed the dynamics of serum metabolic parameters, including glucose, NEFA, TBA, insulin, GH, leptin, and cortisol. Liver transcriptome showed that DLys treatment affected the metabolism-related pathways. Compared with the control group, adipose tissue lipolysis (the adipose triglyceride lipase level was significantly increased), hepatic gluconeogenesis (the G6PC protein level was significantly increased) and fatty acid oxidation (the CPT1A protein level was significantly increased) were promoted in the DLys group. DLys treatment expanded degrees of oxidative phosphorylation in the liver, coming about in a higher NAD+ /NADH proportion and enactment of the SIRT1 signaling pathway. Additionally, the liver protein levels of the DLys group were significantly higher than those of the control group for GHSR, PPAR alpha, and PGC-1. To summarize, inhibition of ghrelin activity can significantly affect metabolism and alter energy levels by enhancing fat mobilization, hepatic fatty acid oxidation and gluconeogenesis without affecting fatty acid uptake and synthesis in the liver.

Feeding Frequency Modulates the Intestinal Transcriptome Without Affecting the Gut Microbiota in Pigs With the Same Daily Feed Intake.

The objective of this study was to elucidate the impacts of irregular eating patterns on gut microbiota and transcriptomic responses in a pig model with different feeding regimens. The experiment involved 24 growing pigs (Duroc × Landrace × Large White, 48 days of age) which were randomly allocated to one of three feeding patterns: one-meal (M1), three-meals (M3), or five-meals (M5) per day with the same daily feed intake. The results showed that different feeding frequencies had no significant effects on the microbial composition of ileal digesta, colonic digesta, colon mucosa, as well as the concentration of SCFAs in colonic digesta. Mucosa transcriptomic profiling data showed the pathways related to vitamin metabolism were enriched in the ileum and colon of pigs in the pairwise comparison between M3 and M1 groups. On the other hand, the pathways related to lipid metabolism were enriched in the ileum and colon of pigs in the pairwise comparison between M5 and M1 groups. Lastly, the pathways related to protein metabolism were enriched in the colon in the pairwise comparison between M3 and M1 groups, M5 and M1 groups, M5 and M3 groups, while the ileum was not enriched. Differentially expressed genes (DEG) related to metabolism showed that carbohydrate transport was suppressed in the ileum and enhanced in the colon in M5 and M3 groups compared with the M1 group. Compared with the M3 group, carbohydrate transport in the ileum was enhanced in the M5 group, while in the colon was inhibited. With the increase of feeding frequency, the catabolism, biosynthesis, and transport of lipid in the ileum were suppressed, while those in the colon were enhanced. Compared with the M1 group, amino acid transport in the ileum and colon in the M3 group was enhanced. Amino acid catabolism in the ileum in the M5 group was enhanced compared with M1 and M3 groups. In summary, different feeding frequencies affected the transport of carbohydrate, lipid, and amino acid in the ileum and colon, and affected the catabolism and biosynthesis of lipid in the ileum and colon with a low impact on intestinal microbiota.

Metabolome-Microbiome Responses of Growing Pigs Induced by Time-Restricted Feeding.

Time-restricted feeding (TRF) mode is a potential strategy in improving the health and production of farm animals. However, the effect of TRF on microbiota and their metabolism in the large intestine of the host remains unclear. Therefore, the present study aimed to investigate the responses of microbiome and metabolome induced by TRF based on a growing-pig model. Twelve crossbred growing barrows were randomly allotted into two groups with six replicates (1 pig/pen), namely, the free-access feeding group (FA) and TRF group. Pigs in the FA group were fed free access while the TRF group were fed free access within a regular time three times per day at 07:00-08:00, 12:00-13:00, and 18:00-19:00, respectively. Results showed that the concentrations of NH4-N, putrescine, cadaverine, spermidine, spermine, total biogenic amines, isobutyrate, butyrate, isovalerate, total SCFA, and lactate were increased while the pH value in the colonic digesta and the concentration of acetate was decreased in the TRF group. The Shannon index was significantly increased in the TRF group; however, no significant effects were found in the Fisher index, Simpson index, ACE index, Chao1 index, and observed species between the two groups. In the TRF group, the relative abundances of Prevotella 1 and Eubacterium ruminantium group were significantly increased while the relative abundances of Clostridium sensu sticto 1, Lactobacillus, and Eubacterium coprostanoligenes group were decreased compared with the FA group. PLS-DA analysis revealed an obvious and regular variation between the FA and TRF groups, further pathway enrichment analysis showed that these differential features were mainly enriched in pyrimidine metabolism, nicotinate and nicotinamide metabolism, glycerolipid metabolism, and fructose and mannose metabolism. In addition, Pearson's correlation analysis indicated that the changes in the microbial genera were correlated with the colonic metabolites. In conclusion, these results together indicated that although the overall microbial composition in the colon was not changed, TRF induced the gradient changes of the nutrients and metabolites which were correlated with certain microbial genera including Lactobacillus, Eubacterium_ruminantium group, Eubacterium coprostanoligenes group, Prevotella 1, and Clostridium sensu sticto 1. However, more studies are needed to understand the impacts of TRF on the health and metabolism of growing pigs.