Effect Of H2O2 Treatment On Endothelial Cell Regulation Of Skeletal Muscle Cell Proliferation

Crosstalk communication between endothelial and skeletal muscle cells contributes to cellular function in skeletal muscle. Oxidative stress is a major factor contributing to age-related skeletal muscle dysfunction, increasing mortality, hospitalization rates, and reducing quality of life. Elevated ROS may contribute to a decline in age-related endothelial function, characterized by impaired vasodilation and reduced nitric oxide bioavailability. PURPOSE: To investigate the effect of H2O2 treatment on endothelial cell regulation of skeletal muscle cell proliferation. METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to 100 μM H2O2 for 48 hr. Media was collected from Control (CON) and H2O2-treated cells for use on primary human skeletal muscle satellite cells (hMuSC). hMuSC were grown in a 50:50 mixture of skeletal muscle growth media and HUVEC CON or H2O2 media for 6 days. hMuSC proliferation was measured by EdU (n = 6) and hMuSC viability was assessed by MTT (n = 8). Data are Mean ± SD. RESULTS: Media from H2O2-treated HUVEC lowered the number of EdU+ hMuSC per field of view (CON: 13.4 ± 7.4; H2O2: 10.3 ± 6.2, p < 0.05), but did not alter the percentage (%) of EdU+ cells (CON: 9.6 ± 4.7; H2O2: 8.7 ± 4.7, p > 0.05) compared to CON. There was no difference in MTT between conditions (CON:0.44 ± 0.10; H2O2: 0.43 ± 0.10, p > 0.05). CONCLUSIONS: These preliminary results suggest treatment of endothelial cells with H2O2 may impair skeletal muscle cell proliferation via alterations in endothelial cell-secreted factors. Supported by: This research project was supported by an American Heart Association (AHA) grant 20IPA35360013.