Although the importance of the intestinal microbiota in host growth and health is well known, the relationship between microbiota colonization and muscle development is unclear. In this study, the direct causal effects of the colonization of gut microorganisms on the muscle tissue of piglets were investigated. The body weight and lean mass of germ-free (GF) piglets were approximately 40% lower than those of normal piglets. The deletion of the intestinal microbiota led to weakened muscle function and a reduction in myogenic regulatory proteins, such as MyoG and MyoD, in GF piglets. In addition, the blinded IGF1/AKT/mTOR pathway in GF piglets caused muscle atrophy and autophagy, which were characterized by the high expression of Murf-1 and KLF15. Gut microbiota introduced to GF piglets via fecal microbiota transplantation not only colonized the gut but also partially restored muscle growth and development. Furthermore, the proportion of slow-twitch muscle fibers was lower in the muscle of GF piglets, which was caused by the reduced short-chain fatty acid content in the circulation and impaired mitochondrial function in muscle. Collectively, these findings suggest that the growth, development and function of skeletal muscle in animals are mediated by the intestinal microbiota.
The change characteristics of intestinal microbial succession and the correlation with the production of two important types of bacterial metabolites (short chain fatty acids and bioamine) in piglets during the early stage were fully explored in this study. Six piglets from different litters with the same birth time were selected, weighted and euthanized at 1, 7, 14, 21, 28, 35, and 42 days of age. During this stage, the piglets grew quickly with gradual increases in blood levels of growth hormone and insulin, and in the intestinal developmental index and immunity. 16s rRNA analysis indicated the alpha diversity of colonic microbiome community was higher than ileum. However, the composition change in the ileal microbiota was more dramatic over time. Lactobacillus genus was the dominant bacteria in piglets' ileum while Prevotella and Ruminococcaceae genera were the dominant bacteria in colon up to weaning. Gut bacterial community of the piglets showed obvious differences between the three different phases: newborn, before weaning, and post weaning. This was similar to the morphological change pattern of pigs' gut. Total SCFA content in the colon of pigs showed almost a 20-fold increase at day 42 compared to the value at day 1. The percentage of acetic acid among the total SCFAs dropped quickly from 74.5% at day 1 to 36.5% at day 42, while butyric acid and propionic acid showed significant increases at the stage. The histamine level increased and putrescine level decreased markedly in the colon with time while the amounts of total bioamines, tyramine and spermidine were devoid of changes. Dozens bacteria taxa showed highly correlations with SCFAs and bioamines. These findings provide an expanded view of the dynamic pig gut and gut microbiome at the important early growth stage.
In the present study we have examined whether p38 mitogen activated protein kinase (p38 MAPK) signal pathway interacts with calcium signal on lipid accumulation in primary preadipocytes of mice. The primary preadipocytes were treated with p38 MAPK inhibitor SB203580, blockers and excitomotors of calcium channel for 24 h, respectively. Intracellular triglyceride (TG) content was measured by triglyceride kit and lipid accumulation was determined by Oil Red O staining. Meanwhile, the mRNA expressions of peroxisome proliferators-activated receptor gamma (PPARγ) gene, fatty acid synthetase (FAS) gene, lipoprotein lipase (LPL) gene, vitamin D receptor (VDR) gene and extracellular Ca(2+)-sensing receptor (CaSR) gene were analyzed with real-time PCR. The protein content and phosphorylation of VDR and p38 were tested with Western Blotting. The data showed that intracellular TG content and the mRNA expression levels of PPARγ, FAS, LPL in N group and L group as well as FAS, LPL in C group were increased significantly (P < 0.01) compared to the control. On the contrary, intracellular TG content and the mRNA expression levels of PPARγ, FAS in B group as well as intracellular TG content and PPARγ, FAS, LPL in SB group and B+SB group were decreased significantly (P < 0.01). VDR mRNA expression and protein content were decreased in B, C, and SB added groups (P < 0.01). In addition, p38 phosphorylation levels increased in N and L groups (P < 0.01) and decreased in SB added groups (P < 0.01). These findings suggest that p38 MAPK pathway through regulating VDR mRNA expression participates in mediation of calcium signal and affects calcium signal regulating lipid accumulation in mice preadipocytes through changing PPARγ, FAS and LPL mRNA expression. In addition, calcium signal have a feedback effect in phosphorylation of p38.
This study investigated the effect of Lactobacillus plantarum strain 299v on gut health in suckling piglets. Sixty newborn piglets were assigned to control and probiotic treatments, with three litters per treatment (ten piglets/litter). From days 1 to 20 of life, piglets were orally administered a placebo of 0.1% peptone or 1.0 × 1010 CFU L. plantarum 299v daily. Six piglets per treatment were sacrificed on day 20, and intestinal tissues (including duodenum, jejunum, ileum and colon) and the intestinal contents from colon segments were collected. The results demonstrated that piglets treated with L. plantarum 299v had a lower diarrhoea incidence than the controls. L. plantarum 299v administration significantly increased the ratio of the villus height to the crypt depth in the jejunum and ileum, as well as the mRNA expression of jejunal occludin and ileal zonula occludens 1 (ZO‐1). The L. plantarum treatment also increased the mRNA abundance of porcine β‐defensin 2 (pBD2) and pBD3 in the jejunum and ileum and of toll‐like receptors (TLRs), such as TLR2, TLR4, TLR6 and TLR9 in the ileum, and significantly upregulated the mRNA abundances of ileal pBD1 and colonic TLR4. Additionally, the L. plantarum 299v treatment significantly changed the structure of the colonic microbiota, as evidenced by the obvious increases in the relative abundances of the phyla Firmicutes and Actinobacteria and of the genus Lactobacillus. Our findings indicate that L. plantarum 299v facilitates the gut health of suckling piglets, probably by improving the intestinal morphology and intestinal barrier function and by modifying the structure of the gut microbiota.
FATP1 plays an important role in the trafficking of free fatty acids in adipocytes, however, its precise function and relationship with other fatty acid transporters all remain poorly understood. In this study, FATP1 gene silencing was induced by transfecting siRNA of target sequence into chicken preadipocytes, then the expression of FABP was found down-regulated while the expression of FAT was raised. In addition, differential inhibition of the cells was observed and the expressions of PPARγ and C/EBPα were found down-regulated. Moreover, the silencing also induced the down-regulation of FAS and inhibited the adipogenesis in adipocytes. Of specific interest here was that FATP1 silencing significantly improved the expressions and activities of cell apoptotic factors Caspases 3 and BCL2 associated X protein (Bax). Consequently, FATP1 deficiency prevented the differentiation while induced apoptosis in chicken preadipocytes.
Clostridium sporogenes (C. sporogenes), as a potential probiotic, metabolizes tryptophan and produces an anti-inflammatory metabolite, indole-3-propionic acid (IPA). Herein, we studied the effects of C. sporogenes and its bioactive metabolite, IPA, on skeletal muscle development and chronic inflammation in mice. In the in vivo study, the muscle tissues and serum samples of mice with C. sporogenes supplementation were used to analyze the effects of C. sporogenes on muscle metabolism; the IPA content was determined by metabonomics and ELISA. In an in vitro study, C2C12 cells were exposed to lipopolysaccharide (LPS) alone or LPS + IPA to verify the effect of IPA on muscle cell inflammation by transcriptome, and the involved mechanism was revealed by different functional assays. We observed that C. sporogenes colonization significantly increased the body weight and muscle weight gain, as well as the myogenic regulatory factors’ (MRFs) expression. In addition, C. sporogenes significantly improved host IPA content and decreased pro-inflammatory cytokine levels in the muscle tissue of mice. Subsequently, we confirmed that IPA promoted C2C12 cells’ proliferation by activating MRF signaling. IPA also effectively protected against LPS-induced C2C12 cells inflammation by activating Pregnane X Receptor and restoring the inhibited miR-26a-2-3p expression. miR-26a-2-3p serves as a novel muscle inflammation regulatory factor that could directly bind to the 3′-UTR of IL-1β, a key initiator factor in inflammation. The results suggested that C. sporogenes with its functional metabolite IPA not only helps muscle growth development, but also protects against inflammation, partly by the IPA/ miR-26a-2-3p /IL-1β cascade.
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