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Effects of in Vitro Exercise‐Induced Endothelial Shear Stress on Oxidative Stress and Vasoconstriction Gene Expression
Author(s) -
Garcia Mario,
Morales-Acuna Francisco,
Conde Daniel,
Gurovich Alvaro N.
Publication year - 2020
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.2020.34.s1.06168
Subject(s) - sod2 , oxidative stress , endothelial dysfunction , pulsatile flow , endothelin 1 , endocrinology , vasoconstriction , vasodilation , medicine , endothelium , in vivo , endothelin receptor , gene expression , aerobic exercise , superoxide dismutase , chemistry , biology , gene , biochemistry , receptor , genetics
Background Sedentary lifestyle contributes to endothelial dysfunction through increasing oxidative stress and endothelin‐1, a potent vasoconstrictor that contributes to the pathogenesis of vascular disease. Exercise‐induced endothelial shear stress (ESS) elicits a mechanical stimulus leading to intracellular signaling transduction regulating cellular function that might protect against endothelial dysfunction through promotion of the expression of anti‐oxidative and pro‐vasodilatory genes. Although it has been reported that increments on steady ESS has led to improvements in endothelial function in vivo , the role of pulsatile exercise induced ESS and varying exercise intensities in vitro has not been explored yet. Therefore, the purpose of this study was to determine the effects of pulsatile exercise‐induced ESS based on in vivo exercise intensity on anti‐oxidative (superoxide dismutase, SOD2) and vasoconstrictive (endothelin 1, EDN‐1) gene expression. Methods Commercially available human carotid artery endothelial cells (Sigma‐Aldrich Comp.) were cultured until 95–100% confluence and exposed to pulsatile resting ESS (18 dynes/cm 2 ) for 5 hours, followed by 1 hour at resting ESS, low‐intensity exercise‐induced ESS (35 dynes/cm 2 ), or high‐intensity exercise‐induced ESS (80 dynes/cm 2 ) on parallel fluidic units (Ibidi USA, Inc.). Ibidi pump software (PumpControl Software 1.5.4) was used to control shear stress intensity. Total RNA was extracted and reverse transcribed followed by measurement of EDN‐1 and SOD2 mRNA expression via qRT‐PCR. qRT‐PCRs were performed in duplicates and changes in gene expression were calculated using the ΔΔ‐CT method with GAPDH used as the normalizing control gene. Statistical analysis was performed using graphpad prism 8 software (Graphpad Software.). Significance was considered at p<0.05. Results mRNA expression of SOD2 increased 2.5‐fold after exposure to low‐intensity exercise‐induced ESS (p<0.05). EDN‐1 showed a 5‐fold increase following exposure to low‐intensity exercise‐induced ESS (p<0.05) in comparison to resting conditions. High‐intensity pulsatile exercise‐induced ESS decreased the expression of EDN‐1 in comparison to resting and low‐intensity conditions (p<0.05). Conclusion Pulsatile exercise‐induced ESS improved anti‐oxidative properties in human carotid artery endothelial cells. It seems that higher exercise intensities are required to downregulate the expression of vasoconstrictive genes.