Departments of Obstetrics & Gynecology, Wayne State University School of Medicine, Detroit, Michigan.
Anatomy& Cell Biology, Wayne State University School of Medicine, Detroit, Michigan.
Alcohol Clin Exp Res. 2018 Jan;42(1):53-60. doi: 10.1111/acer.13534. Epub 2017 Nov 22.
Maternal alcohol abuse leading to fetal alcohol spectrum disorder (FASD) includes fetal growth restriction (FGR). Ethanol (EtOH) induces apoptosis of human placental trophoblast cells, possibly disrupting placentation and contributing to FGR in FASD. EtOH facilitates apoptosis in several embryonic tissues, including human trophoblasts, by raising intracellular Ca . We previously found that acute EtOH exposure increases trophoblast apoptosis due to signaling from both intracellular and extracellular Ca . Therefore, nifedipine, a Ca channel blocker that is commonly administered to treat preeclampsia and preterm labor, was evaluated for cytoprotective properties in trophoblast cells exposed to alcohol.
Human first-trimester chorionic villous explants and the human trophoblast cell line HTR-8/SVneo (HTR) were pretreated with 12.5 to 50 nM of the Ca channel blocker nifedipine for 1 hour before exposure to 50 mM EtOH for an additional hour. Intracellular Ca concentrations were monitored in real time by epifluorescence microscopy, using fluo-4-AM. Apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), accumulation of cytoplasmic cytochrome c, and cleavage rates of caspase 3 and caspase 9.
The increase in intracellular Ca upon exposure to EtOH in both villous explants and HTR cells was completely blocked (p < 0.05) when pretreated with nifedipine, accompanied by inhibition of EtOH-induced release of cytochrome c, caspase activities, and TUNEL.
This study indicates that nifedipine can interrupt the apoptotic pathway downstream of EtOH exposure and could provide a novel strategy for future interventions in women with fetuses at risk for FASD.
导致胎儿酒精谱系障碍（FASD）的母体酒精滥用包括胎儿生长受限（FGR）。乙醇（EtOH）诱导人胎盘滋养层细胞凋亡，可能破坏胎盘形成并导致 FASD 中的 FGR。乙醇通过提高细胞内 Ca ，促进几种胚胎组织包括人滋养层细胞的凋亡。我们之前发现，急性 EtOH 暴露会增加滋养层细胞的凋亡，这是由于细胞内和细胞外 Ca 的信号传递。因此，硝苯地平是一种常用于治疗子痫前期和早产的钙通道阻滞剂，用于评估其对暴露于酒精的滋养层细胞的细胞保护特性。
人类早孕绒毛绒毛外植体和人滋养层细胞系 HTR-8/SVneo（HTR）在暴露于 50mM EtOH 前用 12.5 至 50nM 的钙通道阻滞剂硝苯地平预处理 1 小时。通过使用 fluo-4-AM 实时荧光显微镜监测细胞内 Ca 浓度。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）、细胞质细胞色素 c 的积累和 caspase 3 和 caspase 9 的切割率评估细胞凋亡。
硝苯地平预处理完全阻断了绒毛外植体和 HTR 细胞暴露于 EtOH 后细胞内 Ca 的增加（p < 0.05），同时抑制了 EtOH 诱导的细胞色素 c 释放、caspase 活性和 TUNEL。
本研究表明，硝苯地平可以阻断 EtOH 暴露后的凋亡途径，为未来对有 FASD 风险胎儿的妇女进行干预提供了新策略。
