Abstract
While triclosan (TCS) exerts detrimental effects on female reproduction, the effect of TCS-derived toxins on porcine oocytes during in vitro maturation (IVM) is unclear. This study investigated the effects of TCS on mitochondrion-derived reactive oxygen species (ROS) production and apoptosis pathways during porcine oocyte maturation. Porcine oocytes were treated with TCS (1, 10, and 100 μM) and triphenylphosphonium chloride (Mito-TEMPO; 0.1 μM), and matured cumulus oocyte complexes (COCs) were stained with orcein, dichlorofluorescein diacetate (DCF-DA), and Mito-SOX. Proteins and mRNA levels of factors related to cumulus expansion and mitochondrion-mediated apoptosis and antioxidant enzymes were analyzed by western blotting and reverse-transcription polymerase chain reaction (RT-PCR), respectively. Meiotic maturation and cumulus cell expansion significantly decreased for COCs after TCS treatment along with an increase in mitochondrial superoxide levels at 44 h of IVM. Further, mitochondrion-related antioxidant enzymes and apoptosis markers were significantly elevated in porcine COCs following TCS-mediated oxidative damage. The protective effect of Mito-TEMPO as a specific superoxide scavenger from TCS toxin improved the maturation capacity of porcine COCs. Mito-TEMPO downregulated the mitochondrial apoptosis of TCS-exposed porcine COCs by reducing superoxide level. In conclusion, our data demonstrate that TCS mediates toxicity during porcine oocyte maturation through superoxide production and mitochondrion-mediated apoptosis.
Highlights
Triclosan (TCS) is a broad-spectrum antimicrobial agent widely used in pharmaceuticals and personal-care products at concentrations of 0.1–0.3% (w/w) [1]
Changes in the morphology of cumulus cells significantly reduced (p < 0.001) following exposure to 100 μM TCS as compared with that reported for non-treated matured cumulus oocyte complexes (COCs). mRNA levels of the factors related to cumulus cell expansion (Ptx3, Tnfaip6, and Has2) were significantly lower in porcine-matured COCs treated with 10 and 100 μM TCS (p < 0.01 for Has2 and PTX3 and p < 0.001 for Tnfaip6; Figure 1b) than in non-treated COCs
To evaluate the protective effects of Mito-TEMPO against TCS-induced impairment of porcine oocyte maturation, we investigated changes in meiotic maturation and cumulus cell expansion in Mito-TEMPO-treated COCs subjected to TCS pretreatment
Summary
Triclosan (TCS) is a broad-spectrum antimicrobial agent widely used in pharmaceuticals and personal-care products at concentrations of 0.1–0.3% (w/w) [1]. TCS is thought to exert potential biochemical and genetic toxicities in Eisenia fetida [8] It stimulates the release of superoxide radicals from the mitochondrion and interferes with mitochondrial respiration in the human epithelial cell line HaCaT [9]. These responses following TCS exposure may cause reproductive defects and embryo implantation failure in females. Our previous study demonstrated the important role of BPA-induced ROS and superoxide production in mitochondrial functions and mitochondrion-mediated apoptosis activation during in vitro porcine oocyte maturation [16]. We determined the protective effects of triphenylphosphonium chloride (Mito-TEMPO), a specific superoxide scavenger, against TCS-induced superoxide production by evaluating alterations in mitochondrion-related antioxidant enzymes and apoptosis of porcine COCs
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