Hydroxyurea affects in vitro porcine oocyte maturation through increased apoptosis and oxidative stress

Abstract

Hydroxyurea (HU) is an FDA-approved drug used to treat a variety of diseases, especially malignancies, but is harmful to fertility. We used porcine oocytes as an experimental model to study the effect of HU during oocyte maturation. Exposure of cumulus-oocyte complexes (COCs) to 20 µM (p < 0.01) and 50 µM (p < 0.001) HU reduced oocyte maturation. Exposure to 20 µM HU induced approximately 1.5-fold and 2-fold increases in Caspase-3 (p < 0.001) and P53 (p < 0.01) gene expression levels in cumulus cells, respectively, increased Caspase-3 (p < 0.01) and P53 (p < 0.001) protein expression levels in metaphase II (MII) oocytes and increased the percentage of apoptotic cumulus cells (p < 0.001). In addition, HU decreased the mitochondrial membrane potential (Δφm) (p < 0.01 and p < 0.001) and glutathione (GSH) levels (p < 0.01 and p < 0.001) of both cumulus cells and MII oocytes, while increasing their reactive oxygen species (ROS) levels (p < 0.001). Following parthenogenetic activation of embryos derived from MII oocytes, exposure to 20 µM HU significantly reduced total blastocyst cell numbers (p < 0.001) and increased apoptosis of blastocyst cells (p < 0.001). Moreover, HU exposure reduced the rate of development of 2-cell, 4 -8 cell, blastocyst, and hatching stages after parthenogenetic activation (p < 0.05). Our findings indicate that exposure to 20 µM HU caused significant oxidative stress and apoptosis of MII oocytes during maturation, which affected their developmental ability. These results provide valuable information for safety assessments of HU.

Protective potential of royal jelly against hydroxyurea -induced hepatic injury in rats via antioxidant, anti-inflammatory, and anti-apoptosis properties

“…The current data revealed that HDU triggered significant hepatic inflammation and apoptosis via upregulation of TNF-α and caspase-3 apoptotic pathways expressed by the strong immunostaining of both TNF-α and Caspase-3 proteins liver tissues in HDU treated rats. These latter results are in harmony with previous studies [9] and may be explained by the DNA damage due to increased hepatic NO level [68]. In the current study, the downregulation of TNF-α and Caspase-3 expressions in the liver tissues following oral RJ co-administration reflected the RJ's anti-inflammatory and anti-apoptotic effects against HDU-induced hepatic injury.…”

“…Furthermore, it is recurrently used to treat HIV infections and sickle cell disease, polycythemia vera, and essential thrombocytopenia [1,7,8]. Although its advantageous effect, there are many reports on gonadotoxicity [3,9] cytotoxicity [10], and genotoxicity impact of HDU [11,12]. The integrated mechanism of HDU-induced gonadotoxicity (decreased sperm production and spermatogenic arrest, and reduced oocyte maturation) and cytotoxicity is explained by the overproduction of the formation of the reactive oxygen species [3,9].…”

“…Although its advantageous effect, there are many reports on gonadotoxicity [3,9] cytotoxicity [10], and genotoxicity impact of HDU [11,12]. The integrated mechanism of HDU-induced gonadotoxicity (decreased sperm production and spermatogenic arrest, and reduced oocyte maturation) and cytotoxicity is explained by the overproduction of the formation of the reactive oxygen species [3,9]. Also, HDU induced apoptosis in fetal tissues [13,14] and cancer cell lines [15,16].…”

Circular RNA LONP2 regulates proliferation, invasion, and apoptosis of bladder cancer cells by sponging microRNA-584-5p

“…The detection of apoptosis was assessed using FITC Annexin V Apoptosis Detection Kit (Biolegend, California, USA), as previously described [ 27 ]. T24 and 5637 cells after different treatment were resuspended in Annexin V binding buffer at a concentration of 1.0 × 10 7 cells/mL.…”

RUNX1 (RUNX family transcription factor 1), a target of microRNA miR-128-3p, promotes temozolomide resistance in glioblastoma multiform by upregulating multidrug resistance-associated protein 1 (MRP1)

“…The percentage of apoptotic cells was detected as previously described [ 30 ]. U87MG and A172 cells were trypsinized and resuspended in Annexin V binding buffer (FITC-Annexin V Apoptosis Detection Kit with 7-AAD, Biolegend, California, USA) at a concentration of 1.0 × 10 6 cells/mL.…”