Urotensin II Inhibits Doxorubicin-Induced Human Umbilical Vein Endothelial Cell Death by Modulating ATF Expression and via the ERK and Akt Pathway

Abstract

Background and PurposeRegulation of the homeostasis of vascular endothelium is critical for the processes of vascular remodeling and angiogenesis under physiological and pathological conditions. Urotensin II (U-II), a potent vasoactive peptide, participates in vascular and myocardial remodeling after injury. We investigated the protective effect of U-II on doxorubicin (DOX)-induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs) and the potential mechanisms involved in this process.Experimental ApproachCultured HUVECs were treated with vehicle, DOX (1 µM), U-II, or U-II plus DOX. Apoptosis was evaluated by DNA strand break level with TdT-mediated dUTP nick-end labeling (TUNEL) staining. Western blot analysis was employed to determine the related protein expression and flow cytometry assay was used to determine the TUNEL positive cells.Key ResultsU-II reduced the quantity of cleaved caspase-3 and cytosol cytochrome c and increased Bcl-2 expression, which results in protecting HUVECs from DOX-induced apoptosis. U-II induced Activating transcription factor 3 (ATF3) at both mRNA and protein levels in U-II-treated cells. Knockdown of ATF3 with ATF3 siRNA significantly reduced ATF3 protein levels and U-II protective effect under DOX-treated condition. U-II downregulated p53 expression in DOX-induced HUVECs apoptosis, and it rapidly activated extracellular signal-regulated protein kinase (ERK) and Akt. The DOX induced change of p53 was not affected by U-II antagonist (urantide) under ATF-3 knockdown. The inhibitory effect of U-II on DOX-increased apoptosis was attenuated by inhibitors of ERK (U0126) and PI3K/Akt (LY294002).Conclusion and ImplicationsOur observations provide evidence that U-II protects HUVECs from DOX-induced apoptosis. ERK-Akt phosphorylation, ATF3 activation, and p53 downregulation may play a signal-transduction role in this process.

Chemotaxis-driven assembly of endothelial barrier in a tumor-on-a-chip platform

“…In addition to cancer spheroids, the drug was also found to be cytotoxic to HUVECs cells as the endothelial barrier was found to have disappeared upon exposure to Doxorubicin. This finding is consistent with other reports, which showed the cytotoxic effect of Doxorubicin on endothelial cells 37 .…”

Urotensin II activates the ferroptosis pathway through circ0004372/miR-124/SERTAD4 to promote the activation of vascular adventitial fibroblasts

“…Zhang et al (2022) also found that Ang II increased the proliferation and ferroptosis levels of cardiac microvascular endothelial cells at the same time. In addition, UII did not affect AF apoptosis (data not shown), which was consistent with the results of Chen et al (2014). Chen et al (2014) showed that UII did not affect the apoptosis of human umbilical vein endothelial cells, but inhibited doxorubicininduced cell apoptosis.…”

Exposure to Dengue Envelope Protein Domain III Induces Nlrp3 Inflammasome-Dependent Endothelial Dysfunction and Hemorrhage in Mice

“…In line with the data on IL-1β, TNF-α, and thrombomodulin ( Figure 1 and Figures S1–S3 ), DENV and rEIII treatment induced endothelial cell death in a dose-dependent manner ( Figure 2A ). Overall, various cell death inducers, including doxorubicin (apoptosis) ( 36 , 37 ), rapamycin (autophagy) ( 38 ), erastin (ferroptosis) ( 39 ), TNF-α (necroptosis) ( 40 , 41 ), nigericin (pyroptosis) ( 42 ), served as positive control agents to induce respective cell death pathway of the tested endothelial cells ( Figures 2B, C , dead cell population adjusted to 100%; Figure S3 , flow cytometry gating and calculation). Notably, when compared with cell death agonists, DENV and rEIII treatment induced considerable pyroptosis, necroptosis, and ferroptosis responses in the endothelial cells, but only minor manifestations of apoptosis ( Figure 2B , % of total cells; 2C, % of total dead cell).…”