Zearalenone Induces Endothelial Cell Apoptosis through Activation of a Cytosolic Ca2+/ERK1/2/p53/Caspase 3 Signaling Pathway

Abstract

Zearalenone (ZEN) is a mycotoxin that has been reported to damage various types of cells/tissues, yet its effects on endothelial cells (ECs) have never been investigated. Therefore, this study investigates the potential effects of ZEN using bovine aortic ECs (BAECs). In this study, we found that ZEN induced apoptosis of BAECs through increased cleavage of caspase 3 and poly ADP-ribose polymerase (PARP). ZEN also increased phosphorylation of ERK1/2 and p53, and treatment with the ERK1/2 or p53 inhibitor reversed ZEN-induced EC apoptosis. Transfection of BAECs with small interfering RNA against ERK1/2 or p53 revealed ERK1/2 as an upstream target of p53 in ZEN-stimulated apoptosis. ZEN increased the production of reactive oxygen species (ROS), yet treatment with the antioxidant did not prevent EC apoptosis. Similarly, blocking of estrogen receptors by specific inhibitors also did not prevent ZEN-induced apoptosis. Finally, chelation of cytosolic calcium (Ca2+) using BAPTA-AM or inhibition of endoplasmic reticulum (ER) Ca2+ channel using 2-APB reversed ZEN-induced EC apoptosis, but not by inhibiting ER stress using 4-PBA. Together, our findings demonstrate that ZEN induces EC apoptosis through an ERK1/2/p53/caspase 3 signaling pathway activated by Ca2+ release from the ER, and this pathway is independent of ROS production and estrogen receptor activation.

Zearalenone Induces Apoptosis and Autophagy in a Spermatogonia Cell Line

“…In addition, our results showed an increase in the levels of phosphorylated p53 and ERK 1/2 in GC-1 spg cells following ZEN treatment. Lee et al also reported ZEN-induced apoptosis of endothelial cells through an ERK1/2/p53/caspase-3 signaling pathway, which was similar to our results [38]. ERK activation mediates apoptosis following DNA damage, and p53 is phosphorylated on multiple residues in the amino-and carboxy-terminal domains by several different protein kinases when exposed to DNA damage stress.…”

The Influence of Zearalenone on Selected Hemostatic Parameters in Sexually Immature Gilts

“…The main limitations of this study were its small sample size and the fact that only selected hemostatic parameters were analyzed. Although these limitations can reduce statistical power to detect differences between the experimental groups, we believe that this study provides new information, making it a valuable contribution to the latest in vitro research into ZEN’s effect on endothelial cells and the vascular system [ 5 , 40 , 53 , 54 ]. The present findings suggest that prolonged exposure to ZEN, even at doses below NOAEL values, can affect the porcine vascular system.…”

“…Kitambi et al [ 39 ] demonstrated that ZEN induced selective changes only in the intraocular vasculature of zebrafish. In a study by Lee et al [ 40 ], ZEN (30 µM) stimulated apoptosis in bovine aortic endothelial cells independently of ERs or ROS production. According to a recent study by Güner [ 5 ], ZEN causes vascular irritation in a dose-dependent manner.…”

Introduction to the Toxins’ Special Issue on Evaluation of Cytotoxicity and Cytoprotection Effects of Natural Toxins

“…Studies of mycotoxins in animal cell lines are here presented for two of these cells against ZEN. Two studies in different cell lines are presented: in chicken granulosa cells and bovine aortic ECs (BAECs) [5,6]. The study in chicken granulosa cells was carried out by Zhu et al [5] and was focused on studying the effect on the function of apoptosis and autophagy by gene expression (Bax, Bcl-2, Cyt C, and Caspase-9 and -3 for apoptosis; and LC3-II and Beclin-1 for autophagy).…”

“…This cemented an understanding on the signaling pathway for autophagy activated by ZEN [5]. In bovine aortic ECs cells [6], a wide signaling pathway for ZEN was studied in apoptosis through caspase-3 and PARP (poly ADPribose polymerase) (ERK1/2/p53/caspase-3), but was independent of ROS production and estrogen receptor activation associated with ZEN.…”