Jiang Bo, Zhang Zhiqun, Zheng Pengfei, Ju Li, Tang Kai, Lou Yue
Department of Pediatric Orthopaedics, Nanjing Children's Hospital Affiliated to Nanjing Medical University Nanjing 210008, China.
Int J Clin Exp Pathol. 2014 Jan 15;7(2):677-84. eCollection 2014.
To investigate the apoptotic gene expression of placenta in an all-trans-retinoic acid (ATRA) induced fetus congenital clubfoot pregnant rat model.
Sprague-Dawley (SD) rats were divided randomly into ATRA-exposed group and control group. On day 10 of pregnancy, a dose of 120 mg/kg ATRA dissolved in mineral oil was given intragastrically to the rats in the ATRA-exposed group and equivalent volume of mineral oil was given intragastrically to the control rats. Fetuses were delivered on day 20 of pregnancy, the placenta was collected for the pathological and biochemical analysis.
Clubfoot-like deformity fetuses were observed in the ATRA-exposed group and none with deformity was found in the control group. The pro-apoptosis in placenta of ATRA-exposed group was measured by flow cytometry. Moreover, compared with the control group, lower expression of Bcl-2 and higher expression of BAX were found in the ATRA-exposed group in both mRNA and protein level. Immunohistochemical labeling of Bcl-2 in the control group was more intense while BAX labeling in the ATRA-exposed group was more intense. Additionally, the caspase-3 activity was also significantly increased in the ATRA-exposed group than control group.
In our research, we found a pro-apoptosis in placenta in the ATRA-exposed pregnant rats, indicating a possible association between placental apoptosis and congenital clubfoot.
研究全反式维甲酸(ATRA)诱导的胎儿先天性马蹄内翻足孕鼠模型中胎盘凋亡基因的表达情况。
将Sprague-Dawley(SD)大鼠随机分为ATRA暴露组和对照组。妊娠第10天,给ATRA暴露组大鼠灌胃120 mg/kg溶于矿物油的ATRA,给对照组大鼠灌胃等量体积的矿物油。妊娠第20天分娩胎儿,收集胎盘进行病理和生化分析。
ATRA暴露组观察到类似马蹄内翻足畸形的胎儿,对照组未发现畸形胎儿。采用流式细胞术检测ATRA暴露组胎盘的促凋亡情况。此外,与对照组相比,ATRA暴露组在mRNA和蛋白水平上均发现Bcl-2表达降低,BAX表达升高。对照组Bcl-2的免疫组化染色更强,而ATRA暴露组BAX染色更强。此外,ATRA暴露组的caspase-3活性也比对照组显著增加。
在本研究中,我们发现ATRA暴露的孕鼠胎盘中存在促凋亡现象,提示胎盘凋亡与先天性马蹄内翻足之间可能存在关联。