Abstract
2-Bromopropane (2-BP) is used as an alternative to ozone-depleting cleaning solvents. Previously, we reported that 2-BP has cytotoxic effects on mouse blastocysts and is associated with defects in subsequent development. In the present work, we show that 2-BP induces apoptosis in the inner cell mass of mouse blastocysts, and inhibits cell proliferation. Both effects are suppressed by resveratrol, a grape-derived phytoalexin with known antioxidant and anti-inflammatory properties. 2-BP-treated blastocysts displayed lower levels of implantation (compared to controls) when plated on culture dishes in vitro, and a reduced ability to proceed to later stages of embryonic development. Pretreatment with resveratrol prevented 2-BP-induced disruption of embryonic development, both in vitro and in vivo. Further investigation of these processes revealed that 2-BP directly promotes ROS generation, loss of mitochondrial membrane potential (MMP), and activation of caspase-3, whereas resveratrol effectively blocks 2-BP-induced ROS production and the accompanying apoptotic biochemical changes. Our results collectively imply that 2-BP triggers the mitochondrion-dependent apoptotic pathway via ROS generation, and the antioxidant activity of resveratrol prevents 2-BP-induced toxicity.
Highlights
2-Bromopropane (2-BP) is used as an alternative to ozone-depleting cleaning solvents
We further examined the effects of resveratrol on 2-BP-promoted cell death in mouse blastocysts. 2-BP-induced apoptosis was prevented by pretreatment with 10–20 μM resveratrol (Figure 1B)
Differential staining followed by cell counting was used to examine cell proliferation within blastocysts to explore the protective effects of resveratrol on cells treated with 2-BP (5 μM) for 24 h
Summary
2-Bromopropane (2-BP), a cleaning agent, is used as an alternative to ozone-depleting solvents. Our recent study results indicate that 2-BP induces developmental injury via induction of cell apoptosis processes in oocyte maturation and early-stage embryos [14]. These findings clearly suggest that short-term exposure to 2-BP is a risk factor for normal mouse embryonic development, and may inhibit oocyte maturation in infertile subjects. Our results indicated that resveratrol reduces ethanol-induced injury (including apoptosis), inhibition of cell proliferation, and retardation of embryonic development, because of the antioxidant properties of the material [28]. Pretreatment with resveratrol effectively suppressed 2-BP-induced injury, including cell apoptosis, inhibition of cell proliferation, and retardation of embryonic development, both in vitro and in vivo. Further studies were conducted to elucidate the mechanisms underlying these effects
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