DNA甲基化转移酶1与2,3,7,8-四氯二苯并对二噁英诱导小鼠腭裂的相关性研究

doi:10.13701/j.cnki.kqyxyj.2019.02.007

口腔医学研究 ›› 2019, Vol. 35 ›› Issue (2): 129-132.DOI: 10.13701/j.cnki.kqyxyj.2019.02.007

DNA甲基化转移酶1与2,3,7,8-四氯二苯并对二噁英诱导小鼠腭裂的相关性研究

李勇¹,张伟²,安莉¹,宋红权¹,张兴伟¹,高雨蔚¹,臧棨光¹,焦晓辉^1*

1. 哈尔滨医科大学附属口腔医院口腔颌面外科黑龙江哈尔滨 150001;
2. 哈尔滨医科大学附属第四医院口腔科黑龙江哈尔滨 150001

收稿日期:2018-08-15 出版日期:2019-02-18 发布日期:2019-02-25
通讯作者: 焦晓辉,E-mail:jiaoxiaohuidoctor@163.com
作者简介:李勇(1993~ ),男,河北省保定人,硕士在读,研究方向为唇腭裂发病机制及诊断等。
基金资助:
国家重点研究计划(编号:2016YFC1000504)

Correlation between Methyltransferase DNMT1 and TCDD-induced Cleft Palate in Mice

LI Yong¹, ZHANG Wei², AN Li¹, SONG Hong-quan¹, ZHANG Xing-wei¹, GAO Yu-wei¹, ZANG Qi-guang¹, JIAO Xiao-hui^1*

1. Department of Oral and Maxillofacial Surgery, Affiliated Stomatological Hospital of Harbin Medical University, Harbin 150001, China;
2. Department of Stomatology, Fourth Affiliated Hospital of Harbin Medical University, Harbin 150001, China.

Received:2018-08-15 Online:2019-02-18 Published:2019-02-25

摘要/Abstract

摘要： 目的: 通过检测DNA甲基转移酶1(DNA methyltransferase 1,DNMT1)在四氯二苯并对二噁英(2,3,7,8-tetrachlorodibenzo-paradioxin,TCDD)诱导的腭裂小鼠体内的表达,探讨其与腭裂发病机制的相关性。方法: TCDD通过对腭部中嵴上皮细胞的毒性作用,导致胎鼠腭裂发生。应用TCDD(64 μg/kg)处理胚胎处于第10.5天(E10.5)的C57BL/6J怀孕小鼠建模。分别获取E13.5、E14.5、E15.5和E17.5胎鼠的口腔组织,苏木精-伊红(hematoxylin-eosin staining,HE)染色检测胎鼠腭裂组织。逆转录-定量聚合酶链反应(reverse transcription-quantitative polymerase chain reaction,RT-qPCR)检测以上各组胎鼠DNMT1 mRNA表达水平的变化,Western blot检测DNMT1蛋白表达水平的变化。结果: HE染色结果显示TCDD组腭裂发生率为98.25%,对照组无腭裂发生。RT-qPCR结果显示TCDD组DNMT1 mRNA水平与对照组比较显著增高,差异有统计学意义(P<0.01)。Western blot结果表明与对照组比较,TCDD组DNMT1蛋白表达水平显著增高,差异有统计学意义(P<0.01)。结论: 在TCDD诱导胎鼠腭裂的形成过程中,DNMT1发挥重要的作用。

关键词: 非综合征型唇腭裂, 甲基化, 四氯二苯并对二噁英, DNA甲基转移酶1

Abstract: Objective : To investigate the correlation between DNMT1 and TCDD-induced cleft palate in mice by detecting the expression of DNMT1 in TCDD-induced cleft palate mice. Methods: TCDD could cause fetus rats cleft palate by toxic effects on palatal medial edge epithelial cells. C57BL/6J pregnant mice were treated with TCDD (64 μg/kg) at day 10.5 (E10.5) to be the modeling. Oral tissues of E13.5, E14.5, E15.5, and E17.5 fetal rats were obtained, and HE staining was used to detect the occurrence of fetal cleft palate. Reverse transcription quantitative polymerase chain reaction was used to detect the changes of mRNA expression in DNMT1 of fetal rats. Western blot was used to detect the expression level of DNMT1 protein. Results: HE staining showed that the incidence of cleft palate in group TCDD was 56/57, and there was no cleft palate in control group. RT-qPCR results showed that DNMT1 mRNA in TCDD group was significantly higher than that in control group (P<0.01). Western blot results showed that compared with the control group, the expression level of DNMT1 protein in TCDD group was significantly higher (P<0.01). Conclusion: Methyltransferase DNMT1 plays an important role in the process of TCDD induced cleft palate formation in fetal rats.

Key words: Nonsyndromic cleft, lip with or without cleft palate, Methylation, Tetrachlorodibenzo-P-dioxin, DNA methyltransferase 1

李勇,张伟,安莉,宋红权,张兴伟,高雨蔚,臧棨光,焦晓辉. DNA甲基化转移酶1与2,3,7,8-四氯二苯并对二噁英诱导小鼠腭裂的相关性研究[J]. 口腔医学研究, 2019, 35(2): 129-132.

LI Yong, ZHANG Wei, AN Li, SONG Hong-quan, ZHANG Xing-wei, GAO Yu-wei, ZANG Qi-guang, JIAO Xiao-hui.. Correlation between Methyltransferase DNMT1 and TCDD-induced Cleft Palate in Mice[J]. Journal of Oral Science Research, 2019, 35(2): 129-132.

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DNA甲基化转移酶1与2,3,7,8-四氯二苯并对二噁英诱导小鼠腭裂的相关性研究

Correlation between Methyltransferase DNMT1 and TCDD-induced Cleft Palate in Mice

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