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Skeletal Muscle Extracellular Vesicles Regulate Endothelial Cells in a Fiber Type Dependent Manner
Author(s) -
Kargl Chris,
Yang Autumn,
Jia Zhihao,
Kuang Shihuan,
Shannahan Jonathan,
Gavin Timothy
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.03914
Subject(s) - skeletal muscle , angiogenesis , umbilical vein , myogenesis , gastrocnemius muscle , chemistry , secretion , myocyte , extracellular , extracellular matrix , cd31 , microbiology and biotechnology , biology , biochemistry , endocrinology , medicine , in vitro
Introduction Skeletal muscle regulates endothelial cells (EC) via the secretion of angiogenic signaling factors, including extracellular vesicles (EV). Muscle can be characterized as predominantly oxidative (OXI) or glycolytic (GLY) depending on the expression of specific fiber types. OXI fibers have greater capillary density and secrete greater quantities of several angiogenic factors and EV, however it is unknown if EV signaling is regulated by fiber type. We hypothesized that EV released from OXI muscle (OXI‐EV) induce a greater angiogenic response in cultured EC compared to EV from GLY muscle (GLY‐EV). Methods From male CL57BL/6 mice, OXI muscle was isolated from red portions of the gastrocnemius and soleus; GLY muscle was isolated from white portions of the gastrocnemius and tibialis anterior. EVs were isolated using ultrafiltration‐size exclusion chromatography. EV number and RNA content were measured. Human umbilical vein endothelial cells (HUVEC) were treated with OXI‐EV or GLY‐EV (10 μg/ml) for 24 h; and HUVEC functional angiogenic responses and molecular pathways related to angiogenesis were analyzed. Significance was established at P ≤ 0.05. Results OXI muscle released more EV than GLY muscle (OXI: 8.1x10 6 ; GLY: 4.6x10 6 particles/ml/mg muscle). Selected angiogenic mRNA (vascular endothelial growth factor (VEGF) A and B, angiopoietin (ANG) 1 and 2) and miRNAs (15a, 126, 133a) were not different between OXI‐EV and GLY‐EV, although there was a trend towards greater VEGFA in OXI‐EV. In HUVEC, OXI‐EV promoted greater tube formation (# of tubes: +34%; length: +20%), lowered intracellular superoxide (‐58%), and trended (P < 0.10) toward greater growth (+22%) with no difference in migration compared to GLY‐EVs. Treatment with OXI‐EV promoted greater HUVEC endothelial nitric oxide synthase (eNOS) phosphorylation (+40%) and gene expression (+42%) compared to GLY‐EV. In HUVEC, there was no difference in gene expression of VEGFA, VEGFB, ANG1, or Ang2 between OXI‐EV and GLY‐EV treatments. Conclusions EV from oxidative muscle demonstrate greater angiogenic potential than EV from glycolytic muscle consistent with greater capillarization of oxidative muscle. Thus, greater oxidative vs. glycolytic muscle capillarization is likely regulated in part by the combination of greater release of EV and greater EV angiogenic potential from oxidative muscle fibers.