TCDD诱导C57BL／6J胎鼠腭裂过程中组蛋白H4乙酰化的表达

The expression change of histone H4 acetylation in TCDD-induced cleft palates formation in fetalmice

目的检测在2，3，7，8-四氯二苯二噁英（2，3，7，8-tetrachlorod-ibenzo-P-dioxin，TCDD）诱导的C57BL／6J小鼠腭裂过程中组蛋白H4乙酰化的表达水平，探讨TCDD诱发腭裂与组蛋白H4乙酰化的相关性。方法48只C57BL／6J近交系孕鼠完全随机分为实验组和对照组，在小鼠妊娠第10．5天（gestationday，GD10．5）时，一次性给予孕鼠分别灌服20μg/kgTCDD（实验组）和等剂量玉米油（对照组），分别在GD13．5、GD14．5、GD15．5剖腹取胎鼠头部标本，作冠状位组织切片，采用免疫组织化学染色观察组蛋白H4乙酰化在腭突中的表达，Western Blotting检测胎鼠腭组织组蛋白H4乙酰化的相对表达量。应用SPSS24．0进行统计分析，统计方法采用单因素方差分析和方差同质性检验，方差齐采用Bonferroni检验，方差不齐者采用校正t检验，P〈0．05表示差异有统计学意义。结果组蛋白H4乙酰化主要表达在腭突上皮细胞中，间质细胞少许表达；在腭发育各个主要时期GD13．5、GD14．5、GD15．5腭突上皮细胞的相对表达量分别是：对照组为0．60±0．25、0．92±0．09和0．90±0．09；TCDD组为1．02±0．28、1．61±0．27和1．28±0．13，2组比较差异有统计学意义（P〈0．05）。结论TCDD诱导的C57BL／6J胎鼠腭裂的发生可能与组蛋白H4乙酰化修饰有关。

Objective To evaluate the expression of histone H4 acetylation（Ac-H4） during the cleft palates formation induced by 2,3,7, 8-tetrachlorodibenzo-p-dioxin （TCDD） in C57BL/6J mice. Methods Forty-eight pregnant C57BL/6J mice were completely randomly divided into two groups： TCDD group, mice were treated with 20ug/kg of TCDD on gestation day （GD） 10.5 by gastric perfusion; control group, mice were treated with an equivalent of corn oil. The head samples were collected and sliced in coronal plane on GD13.5, GD14.5 and GD15.5 respectively. Histone H4 acetylation in the palates were evaluated by immunohistochemieal staining and Western Blot in the two groups. Results Histone H4 acetylation was mainly expressed in the palatal epithelial cells and slightly expressed in mesenchymal cells. The expression level of histone H4 acetylation was 0.6002 ±0. 2530, 0. 9180 ± 0. 0941and 0. 8966 ±0. 0908 respectively in control group on GD13.5, GD14.5 and GD15.5 ; while 1. 0229 ± 0. 2779, 1. 6095 ± 0. 2651 and 1. 2758 ± 0. 1251 in TCDD group. There were statistically significant differences between the control group and TCDD group （ P 〈 0. 05 ）. Conclusions The histone H4 acetylation was involved in the cleft palate formation induced by TCDD in C57BL/6J mice.