Journal: International Journal of Molecular Medicine
Article Title: Rifaximin enhances the L-carnitine-mediated preventive effects on skeletal muscle atrophy in cirrhotic rats by modulating the gut-liver-muscle axis
doi: 10.3892/ijmm.2022.5157
Figure Lengend Snippet: Cellular mitochondrial biogenesis in LPS-stimulated rat L6 myocytes. (A) In vitro experimental protocol. (B and C) Effects of L-carnitine and/or rifaximin on the mRNA expression levels of (B) Atrogin-1 and MuRF-1 , and (C) Ppargc1a and Tfam in LPS-stimulated rat L6 myocytes. The mRNA expression levels were measured using reverse transcription-quantitative PCR, and Gapdh was used as an internal control. (D) The mitochondrial content was assessed using quantitative PCR as described in the Materias and methods. (E) Representative images of TMRM live stains corresponding to mitochondrial membrane potential. Scale bar, 50 μ m. (F) Quantification of TMRM intensity per cell; data shown as the mean ± SD for 100 cells per condition in three representative experiments. Nuclei were stained with Hoechst 33342. (G) Measurements of OCR using a seahorse extracellular flux analyzer. (H) Calculations of the basal and maximal respiration rates. Cells were treated with LPS (1.0 μ g/ml) and L-carnitine (5 mM) and/or rifaximin (10 μ M) for 48 h. Quantitative values are indicated as fold changes to the values of non-treated (NT) groups (B, C and D). Data are the mean ± SD; (B, C and D) n=8, (F) n=3, or the mean ± SEM (G and H) n=5. aa P<0.01 vs. LPS (-)/L-CAR (-)/RFX (-), b P<0.05 and bb P<0.01 vs. LPS (+)/L-CAR (-)/RFX (-). LPS, lipopolysaccharide; TMRM, tetramethylrhodamine methyl ester; OCR, oxygen consumption rate; CSAA, choline-sufficient amino acid-defined diet; RFX, rifaximin; L-CAR, L-carnitine; Ppargc1a , peroxisome proliferator-activated receptor γ coactivator-1α; Tfam , mitochondrial transcription factor A; MuRF-1 , muscle RING-finger protein-1.
Article Snippet: L6 rat skeletal muscle myoblasts (cat. no. JCRB9081, Japanese Collection of Research Bioresources Cell Bank) were cultured and differentiated into myotubes as previously described ( , ).
Techniques: In Vitro, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Membrane, Staining
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