Probiotics Ameliorate Gut‐Microbial Dysbiosis, Intestinal Permeability, Systemic Inflammation, and Skeletal Muscle Dysfunction in Cystathionine‐β‐synthase‐Deficient Mice

Recent studies have reported that dietary supplementation of probiotics is a novel approach for enhancing skeletal development and metabolism in both humans and animals. However, the precise role of probiotics in regulating skeletal muscle mass at the cellular and molecular levels in mice still needs to be clarified. Here we show that altered gut‐microbiota eubiosis, systemic inflammation and skeletal muscle dysfunction caused by hydrogen sulfide deficiency and whether the intragastric administration of probiotics (GSL#5) could retard H 2 S deficiency‐related skeletal muscle defects. To test this hypothesis, we employed 8‐week‐old female wild‐type (WT), cystathionine‐beta‐synthase heterozygote knockout (CBS+/−), and CBS+/− mice supplemented with probiotics (GSL#5) at the dosage of 1×10 9 CFU every other day through oral gavage for 6 weeks. In the murine model, H2S deficiency (CBS +/− mice) increased gut permeability, and upregulated the inflammatory cytokines TNFα (TNF), and IL‐17 in the blood and the skeletal muscle. Further, the gut microbiota is significantly altered in CBS‐deficient mice as assessed by the Denaturing gradient gel electrophoresis (DGGE) analysis. We documented that long non‐coding RNA‐PVT1 (lncRNA‐PVT1) is, indeed, upregulated in the soleus muscles of CBS‐deficient mice (4.2 fold), as assessed by RT2 lncRNA qPCR Assay. The mechanistic study shows that depressed H 2 S signaling stimulates the transcription factor C/EBP‐α binding to the promoter of lncRNA‐PVT1 and promotes the expression of lncRNA‐PVT1. Using an electrophoretic mobility shift assay and chromatin immunoprecipitations, we identified that lncRNA‐PVT1 interacts with TNFAIP3 interacting protein 1, TNIP1, leading to the acetylation of NF‐κB (P65 subunit) activity in the soleus muscle of CBS‐deficient mice. Further, activation of P65 region of NF‐κB transcriptionally upregulates atrogin‐1, MuRF‐1 expression. Overexpression of lncRNA‐PVT1 in cultured C2C12 muscle cell lines stimulates an increase in MuRF‐1 (2.3‐fold) and atrogin‐1 (3.7‐fold) expression. In addition, systemic delivery of synthetic lncRNA‐PVT1 in control mice causes the myofibers to undergo atrophy. Gene knockout of lncRNA‐PVT1 in CBS‐deficient mice attenuates atrophy of the myofibers and muscle wasting. Overall, administration of probiotics in CBS‐deficient mice reverses the gut‐microbiota dysbiosis and preserves skeletal muscle mass. In conclusion, our findings provide novel evidence that the administration of probiotics could balance the gut‐microbiota homeostasis as well as skeletal muscle function by modulating systemic inflammation in H 2 S‐deficient mice Support or Funding Information This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .