Hydrogen Sulfide Donor NaHS Improves Metabolism and Reduces Muscle Atrophy in Type 2 Diabetes: Implication for Understanding Sarcopenic Pathophysiology

Figure 3

Diabetic atrophied muscles exhibited a state of heightened oxidative stress (HSOS). (a and b) Superoxide generation was measured in frozen muscle sections of control and diabetic using dihydroethidium-based confocal microscopic staining technique. (c and d) NADPH oxidase in a membrane fraction was assessed according to procedure involving the substrate NADPH and lucigenin chemiluminescence or the Amplex Red/horseradish peroxidase fluorescence-based assays. (e and f) Muscle NADPH oxidase-related isoforms including NOX2 and NOX4 were determined at the mRNA (e) and protein levels (f) using RT-PCR and Western blotting-based techniques. (g) Mitochondrial H₂O₂ generation at the steady state level and in the presence of added glutamate/malate substrates was measured using the Amplex Red/horseradish peroxidase fluorescence-based assay (g). Activities of complexes I (h) and III (i) of the electron transport chain were measured using spectrophotometric-based assay. Abbreviation: C: control; D: diabetic. Values are for at least 6 animals/group. Significantly different from corresponding control values at .

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