Linking In Vitro Models of Endothelial Dysfunction with Cell Senescence

Abstract

Endothelial cell dysfunction is the principal cause of several cardiovascular diseases that are increasing in prevalence, healthcare costs, and mortality. Developing a standardized, representative in vitro model of endothelial cell dysfunction is fundamental to a greater understanding of the pathophysiology, and to aiding the development of novel pharmacological therapies. We subjected human umbilical vein endothelial cells (HUVECs) to different periods of nutrient deprivation or increasing doses of H2O2 to represent starvation or elevated oxidative stress, respectively, to investigate changes in cellular function. Both in vitro cellular models of endothelial cell dysfunction-associated senescence developed in this study, starvation and oxidative stress, were validated by markers of cellular senescence (increase in β-galactosidase activity, and changes in senescence gene markers SIRT1 and P21) and endothelial dysfunction as denoted by reductions in angiogenic and migratory capabilities. HUVECs showed a significant H2O2 concentration-dependent reduction in cell viability (p < 0.0001), and a significant increase in oxidative stress (p < 0.0001). Furthermore, HUVECs subjected to 96 h of starvation, or exposed to concentrations of H2O2 of 400 to 1000 μM resulted in impaired angiogenic and migratory potentials. These models will enable improved physiological studies of endothelial cell dysfunction, and the rapid testing of cellular efficacy and toxicity of future novel therapeutic compounds.

High Glucose Promotes and Aggravates the Senescence and Dysfunction of Vascular Endothelial Cells in Women with Hyperglycemia in Pregnancy

“…First, we found that treating primary HUVECs and cell lines with high glucose increased levels of senescence indicators, including p53, p16, p21 ( Figure 2 C,D), and SA-β-gal ( Figure 2 A,B), which was consistent with what was reported in HIP pregnant women. Then, we constructed two senescence models: replicative senescence and SIPS, because current research has not disclosed the type of senescence observed in HUVECs generated by high glucose [ 20 , 21 ]. We confirmed that the primary HUVEC cells passed down to six generations and the HUVECs treated with 1 mmol/L hydrogen peroxide for 48 h achieved replicative senescence and SIPS, respectively, by analyzing levels of senescence indicators p53, p16, p21 ( Figure 2 C,D), and SA-β-gal ( Figure 2 A,B) in the senescence models.…”

Differential Radiomodulating Action of Olea europaea L. cv. Caiazzana Leaf Extract on Human Normal and Cancer Cells: A Joint Chemical and Radiobiological Approach

“…Pre-treatment with, and irradiation in the presence of, OLC resulted in a marked decrease in the onset of X-ray-induced PS in HUVECs, a widely used model system for endothelium dysfunction [ 43 ] associated with the pro-inflammatory response promoted by the secretome from ectopically senescing cells [ 44 ]. In fact, IR-induced PS is often associated with RT side effects such as tissue fibrosis, organ function disruption, and the elevation of secondary cancer risk as well as of cardiovascular disease incidence [ 45 , 46 ].…”

Endothelial dysfunction due to the inhibition of the synthesis of nitric oxide: Proposal and characterization of an in vitro cellular model

“…In this sense, one can observe that in vitro research is also of great importance for a better understanding of ED and to elucidate the pathways involved in endothelial cells. For these reasons, there is a crescent need for standardization of a method of inducing in vitro endothelial dysfunction ( Aman et al, 2016 ; Gallogly et al, 2021 ; Jimenez Trinidad et al, 2021 ).…”