Cyclophosphamide and acrolein induced oxidative stress leading to deterioration of metaphase II mouse oocyte quality

Abstract

Cyclophosphamide (CTX) is a chemotherapeutic agent widely used to treat ovarian, breast, and hematological cancers as well as autoimmune disorders. Such chemotherapy is associated with reproductive failure and premature ovarian insufficiency. The mechanism by which CTX and/or its main metabolite, acrolein, affect female fertility remains unclear, but it is thought to be caused by an overproduction of reactive oxygen species (ROS). Here, we investigated the effect of CTX on metaphase II mouse oocytes obtained from treated animals (120mg/kg, 24h of single treatment), and oocytes directly exposed to increasing concentrations of CTX and acrolein (n=480; 0, 5, 10, 25, 50, and 100μM) with and without cumulus cells (CCs) for 45min which correlates to the time of maximum peak plasma concentrations after administration. Oocytes were fixed and subjected to indirect immunofluorescence and were scored based on microtubule spindle structure (MT) and chromosomal alignment (CH). Generation of ROS was evaluated using the Cellular Reactive Oxygen Species Detection Assay Kit. Deterioration of oocyte quality was noted when oocytes were obtained from CTX treated mice along with CTX and acrolein treated oocytes in a dose-dependent manner as shown by an increase in poor scores. Acrolein had an impact at a significantly lower level as compared to CTX, plateau at 10μM versus 50μM, respectively. These variation is are associated with the higher amount of ROS generated with acrolein exposure as compared to CTX (p<0.05). Utilization of antioxidant therapy and acrolein scavengers may mitigate the damaging effects of these compounds and help women undergoing such treatment.

Protective Effects of Evening Primrose Oil against Cyclophosphamide-Induced Biochemical, Histopathological, and Genotoxic Alterations in Mice

“…There is evidence in the literature for CP-induced hepatotoxicity [15,16] and some reports of CP-related pancreatic dysfunction and diabetic changes [17,18]. The suggested mechanisms for CP-induced liver toxicity include exposure to the CP metabolite (o-carboxyethyl-phosphoramide mustard), which is related to biliary obstruction [15], oxidative stress (related to another metabolite: acrolein) [5], and lymphocytic infiltration [19]. The CP-induced pancreatic toxicity may be an idiosyncratic reaction or secondary to hepatotoxicity [18].…”

“…Cyclophosphamide is metabolized via the cytochrome P450 system, producing acrolein and phosphoramide mustard [4]. Acrolein is a reactive aldehyde that generates toxic reactive oxygen species (ROS), leading to oxidative stress [5]. Cyclophosphamide toxicity affects many organs, including the liver and pancreas, and is characterized by depletion of glutathione, lipid peroxidation, altered DNA profile, pro-inflammatory response, and apoptosis [6,7].…”

Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation

“…The accumulation of oxidative stress exerts long-term effects on ovarian follicle depletion [27, 50]. In addition, the cyclical production of ROS may cause cumulative damage to DNA that contributes to ovarian aging [51]. Repeated ovarian stimulation has been shown to induce oxidative damage and mitochondrial DNA mutations in ovaries, increase the incidence of oocyte spindle defects, and decrease the quality of oocytes [52-54].…”

HGF and BFGF Secretion by Human Adipose-Derived Stem Cells Improves Ovarian Function During Natural Aging via Activation of the SIRT1/FOXO1 Signaling Pathway

“…The accumulation of oxidative stress has long-term effects on ovarian follicle depletion. In addition, the cyclical production of ROS may cause cumulative damage to DNA that contributes to ovarian aging [33]. Therefore, whether HGF and bFGF play a central role in retarding ovarian aging by reducing ROS needs to be clarified.…”