Impact Of H2O2 Treatment On Primary Human Skeletal Muscle Myotube-induced Endothelial Cell Angiogenesis

PURPOSE: Skeletal muscle atrophy and endothelial cell (EC) dysfunction occur concurrently in aging. Communication between skeletal muscle (HSkM) and the vasculature plays an important role in tissue homeostasis. Factors secreted by cells are critical in cell-to-cell communication, influencing cell survival, metabolism, and regenerative capacity regulation. Factors secreted from H2O2-treated primary human muscle satellite cells impair EC angiogenesis. However, skeletal muscle is comprised primarily of mature myofibers and not satellite cells. It was hypothesized that co-culture of EC with media from H2O2 treated primary human skeletal muscle myotubes would impair angiogenesis. METHODS: Primary HSkM myotubes (n = 9/10) were exposed to 200 μM H2O2 or control (CON) for 48 hr and conditioned media was collected. Human umbilical vein endothelial cells (HUVEC) were treated with HSkM myotube-derived media mixed in a 1:1 ratio with endothelial basal media (EBM) for 6 or 36 hours. HUVEC viability (MTT), proliferation (EdU), migration (scratch test). Paired Student’s T-test was used to identify differences between conditions. Significance: P ≤ 0.05. Data are Mean + SD. RESULTS: Media from H2O2 treated HSkM myotubes did not affect HUVEC viability (CON: 0.41 + 0.04; H2O2: 0.41 + 0.03; p = 0.47), proliferation (CON: 18.9 + 4.0%; H2O2: 20.6 + 3.9%; p = 0.19), migration (CON: 90 + 19 cells; H2O2: 99 + 18 cells; p = 0.09) compared to CON. CONCLUSIONS: Exposure of primary human skeletal muscle myotubes to H2O2 does not impair crosstalk regulation of endothelial cell angiogenesis in contrast to findings from primary human skeletal muscle satellite cells. Supported by: This research project was supported by an American Heart Association (AHA) grant, 20IPA35360013