Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, Taiwan, ROC.
Mol Hum Reprod. 2010 May;16(5):361-72. doi: 10.1093/molehr/gaq004. Epub 2010 Jan 18.
The endocrine disruptor 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been demonstrated to disrupt hormone signalling, reduce fertility, interfere with embryo development and cause spontaneous miscarriage in humans. The precise mechanisms of its effects on early implantation in humans are still unclear. In this study, we examined the relationship between mitochondrial function and dioxin-induced toxicity in JAR cells, a human trophoblast-like cell line. Several experiments were performed to address the effects of TCDD on cell viability, reactive oxygen species (ROS) generation, oxidative damage (indicated by the presence of lipoperoxides and oxidized DNA bases), mitochondrial DNA (mtDNA) copy number, ATP content, mtDNA mutations and the protein levels of p53, Bax, Bcl2, cytochrome c and caspase 3. Increased oxidative damage and mitochondrial dysfunction in TCDD-treated trophoblast-like cells was demonstrated. A 2.58-fold increase in lipid peroxides was detected in cells treated with 2 nM TCDD for 4 h. The oxidative DNA damage marker 8-hydroxy-2'-deoxyguanosine was significantly increased by TCDD treatment in a time-dependent manner. Meanwhile, reductions in mtDNA copy number and ATP content and an increase in mtDNA deletions were found. Furthermore, we observed increased apoptosis, p53 accumulation, Bax overexpression, cytochrome c release and sequential caspase 3 activation after TCDD exposure. These results indicate that oxidative damage and mitochondrial dysfunction may be responsible for the apoptotic effects of TCDD.
内分泌干扰物 2,3,7,8-四氯二苯并对二恶英(TCDD)已被证明可破坏激素信号传递、降低生育能力、干扰胚胎发育并导致人类自然流产。其对人类早期着床的精确作用机制仍不清楚。在这项研究中,我们研究了 JAR 细胞(一种人类滋养细胞样细胞系)中线粒体功能与二恶英毒性之间的关系。进行了几项实验以研究 TCDD 对细胞活力、活性氧(ROS)生成、氧化损伤(由脂过氧化物和氧化 DNA 碱基的存在表示)、线粒体 DNA(mtDNA)拷贝数、ATP 含量、mtDNA 突变以及 p53、Bax、Bcl2、细胞色素 c 和半胱氨酸蛋白酶 3 蛋白水平的影响。研究表明,TCDD 处理的滋养细胞样细胞中氧化损伤和线粒体功能障碍增加。用 2 nM TCDD 处理 4 小时后,检测到细胞中脂过氧化物增加了 2.58 倍。TCDD 处理以时间依赖性方式显著增加氧化 DNA 损伤标志物 8-羟基-2'-脱氧鸟苷。同时,还发现 mtDNA 拷贝数和 ATP 含量减少以及 mtDNA 缺失增加。此外,在 TCDD 暴露后观察到细胞凋亡增加、p53 积累、Bax 过表达、细胞色素 c 释放和顺序激活半胱氨酸蛋白酶 3。这些结果表明,氧化损伤和线粒体功能障碍可能是 TCDD 诱导细胞凋亡的原因。