{"title":"[2,3,7,8-四氯二苯并-对二恶英诱导的胎儿小鼠腭形成过程中整体DNA甲基化的变化]。","authors":"Chen Wang, Xingang Yuan, Yuexian Fu, Shana Zhai","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To investigate global DNA methylation and DNA methyhransferases participation in the mechanism of cleft palate induced by maternal exposure to 2,3,7,8-tetrachlrodibenzo-p-dioxin (TCDD)in mice.</p><p><strong>Methods: </strong>40 pregnant C57BL/6J mice were randomly divided into 2 groups: the control group(n =20) and TCDD-exposure group(n =20).On gestation day 10.5 (GD10.5),the mice in TCDD-group were orally administrated with TCDD 28 μg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13.5,GD14.5,GD15.5,GD16.5,GD17.5,fetal palates were collected for analysis. Global DNA methylation levels were detected by Methylamp TM Global DNA Methylation Quantification Ultra Kit through an ELISA-like reaction. The expression levels of DNA methyltransferases were examined by quantitative real-time PC R(q-PCR).IBM SPSS 20.0 software was applied for statistical analysis. Kolmogorov-Smirnov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P ＜ 0.05 was considered statistically significant.</p><p><strong>Results: </strong>The global DNA methylation level in TCDD-exposure group was significantly higher than that in control group on GD13.5 (49.52％ ±4.03％ vs 33.42％ ± 6.78％,P ＜ 0.01),while lower on GD14.5 (24.10％ ±2.29％ vs 30.12％ ±3.92％,P ＜0.05) and on GD16.5 (32.77％ ±0.98％ vs 36.45％ ± 3.27％,P ＜ 0.05).The expression level of Dnmt1 mRNA in TCDD-exposure group was higher than that in control group on GD13.5(1.28±0.11 vs 1.01 ±0.10,P＜0.05) and on GD16.5(1.04 ±0.05 vs 0.81 ±0.01,P ＜0.01).The expression level of Dnmt3a mRNA in TCDD-exposure group was higher than that in control group on GD13.5 (1.15 ±0.17 vs 0.81 ±0.02,P ＜0.05)and on GD16.5 (1.11 ± 0.06 vs 0.96 ± 0.06,P ＜ 0.05).The expression level of Dnmt3b mRNA in TCDD-exposure group was higher than that in control group on GD14.5(0.97 ±0.06 vs 0.72 ±0.06,P ＜0.01).</p><p><strong>Conclusions: </strong>It is supposed that complicated mechanisms are exist to regulate global DNA methylation levels in palatal tissue of fetal mice. The significant increased DNA methylation level on GD13.5 resulting from up-expression of Dnmt1 and Dnmt3a may be one of the epigenetic mechanisms which cause palate malformation in fetal mice induced by maternal exposure to TCDD.</p>","PeriodicalId":69147,"journal":{"name":"中华整形外科杂志","volume":"32 5","pages":"372-7"},"PeriodicalIF":0.0000,"publicationDate":"2016-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Global DNA methylation changes during palatal formation in fetal mice induced by 2,3,7,8-tetrachlrodibenzo-p-dioxin].\",\"authors\":\"Chen Wang, Xingang Yuan, Yuexian Fu, Shana Zhai\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To investigate global DNA methylation and DNA methyhransferases participation in the mechanism of cleft palate induced by maternal exposure to 2,3,7,8-tetrachlrodibenzo-p-dioxin (TCDD)in mice.</p><p><strong>Methods: </strong>40 pregnant C57BL/6J mice were randomly divided into 2 groups: the control group(n =20) and TCDD-exposure group(n =20).On gestation day 10.5 (GD10.5),the mice in TCDD-group were orally administrated with TCDD 28 μg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13.5,GD14.5,GD15.5,GD16.5,GD17.5,fetal palates were collected for analysis. Global DNA methylation levels were detected by Methylamp TM Global DNA Methylation Quantification Ultra Kit through an ELISA-like reaction. The expression levels of DNA methyltransferases were examined by quantitative real-time PC R(q-PCR).IBM SPSS 20.0 software was applied for statistical analysis. Kolmogorov-Smirnov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P ＜ 0.05 was considered statistically significant.</p><p><strong>Results: </strong>The global DNA methylation level in TCDD-exposure group was significantly higher than that in control group on GD13.5 (49.52％ ±4.03％ vs 33.42％ ± 6.78％,P ＜ 0.01),while lower on GD14.5 (24.10％ ±2.29％ vs 30.12％ ±3.92％,P ＜0.05) and on GD16.5 (32.77％ ±0.98％ vs 36.45％ ± 3.27％,P ＜ 0.05).The expression level of Dnmt1 mRNA in TCDD-exposure group was higher than that in control group on GD13.5(1.28±0.11 vs 1.01 ±0.10,P＜0.05) and on GD16.5(1.04 ±0.05 vs 0.81 ±0.01,P ＜0.01).The expression level of Dnmt3a mRNA in TCDD-exposure group was higher than that in control group on GD13.5 (1.15 ±0.17 vs 0.81 ±0.02,P ＜0.05)and on GD16.5 (1.11 ± 0.06 vs 0.96 ± 0.06,P ＜ 0.05).The expression level of Dnmt3b mRNA in TCDD-exposure group was higher than that in control group on GD14.5(0.97 ±0.06 vs 0.72 ±0.06,P ＜0.01).</p><p><strong>Conclusions: </strong>It is supposed that complicated mechanisms are exist to regulate global DNA methylation levels in palatal tissue of fetal mice. The significant increased DNA methylation level on GD13.5 resulting from up-expression of Dnmt1 and Dnmt3a may be one of the epigenetic mechanisms which cause palate malformation in fetal mice induced by maternal exposure to TCDD.</p>\",\"PeriodicalId\":69147,\"journal\":{\"name\":\"中华整形外科杂志\",\"volume\":\"32 5\",\"pages\":\"372-7\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华整形外科杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华整形外科杂志","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的:探讨母体接触2,3,7,8-四氯二苯并-对二恶英(TCDD)诱发小鼠腭裂的DNA甲基化和DNA甲基转移酶参与机制。方法:将40只C57BL/6J妊娠小鼠随机分为对照组(n =20)和tcdd暴露组(n =20)。妊娠第10.5天(GD10.5)， TCDD组小鼠灌胃TCDD 28 μg/kg，对照组小鼠灌胃等量玉米油。取孕鼠GD13.5、GD14.5、GD15.5、GD16.5、GD17.5处死，收集胎儿腭部进行分析。采用Methylamp TM Global DNA methylation Quantification Ultra Kit，通过elisa样反应检测全局DNA甲基化水平。采用实时荧光定量pcr检测DNA甲基转移酶的表达水平。采用IBM SPSS 20.0软件进行统计分析。采用Kolmogorov-Smirnov检验进行正态分布检验，分布为正态分布。两组间进行独立t检验。P < 0.05为差异有统计学意义。结果:tcdd暴露组GD13.5组整体DNA甲基化水平显著高于对照组(49.52%±4.03% vs 33.42%±6.78%，P < 0.01)， GD14.5组(24.10%±2.29% vs 30.12%±3.92%，P <0.05)和GD16.5组(32.77%±0.98% vs 36.45%±3.27%，P <0.05)。tcdd暴露组GD13.5(1.28±0.11 vs 1.01±0.10,P<0.05)和GD16.5(1.04±0.05 vs 0.81±0.01,P <0.01) Dnmt1 mRNA表达量均高于对照组。tcdd暴露组GD13.5(1.15±0.17 vs 0.81±0.02,P <0.05)和GD16.5(1.11±0.06 vs 0.96±0.06,P <0.05)时Dnmt3a mRNA表达水平均高于对照组。GD14.5时，tcdd暴露组Dnmt3b mRNA表达水平高于对照组(0.97±0.06 vs 0.72±0.06,P <0.01)。结论:胎儿小鼠腭组织DNA甲基化水平可能存在复杂的调控机制。Dnmt1和Dnmt3a基因表达上调导致GD13.5 DNA甲基化水平显著升高，可能是母体接触TCDD导致胎鼠腭畸形的表观遗传机制之一。
[Global DNA methylation changes during palatal formation in fetal mice induced by 2,3,7,8-tetrachlrodibenzo-p-dioxin].
Objective: To investigate global DNA methylation and DNA methyhransferases participation in the mechanism of cleft palate induced by maternal exposure to 2,3,7,8-tetrachlrodibenzo-p-dioxin (TCDD)in mice.
Methods: 40 pregnant C57BL/6J mice were randomly divided into 2 groups: the control group(n =20) and TCDD-exposure group(n =20).On gestation day 10.5 (GD10.5),the mice in TCDD-group were orally administrated with TCDD 28 μg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13.5,GD14.5,GD15.5,GD16.5,GD17.5,fetal palates were collected for analysis. Global DNA methylation levels were detected by Methylamp TM Global DNA Methylation Quantification Ultra Kit through an ELISA-like reaction. The expression levels of DNA methyltransferases were examined by quantitative real-time PC R(q-PCR).IBM SPSS 20.0 software was applied for statistical analysis. Kolmogorov-Smirnov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P ＜ 0.05 was considered statistically significant.
Results: The global DNA methylation level in TCDD-exposure group was significantly higher than that in control group on GD13.5 (49.52％ ±4.03％ vs 33.42％ ± 6.78％,P ＜ 0.01),while lower on GD14.5 (24.10％ ±2.29％ vs 30.12％ ±3.92％,P ＜0.05) and on GD16.5 (32.77％ ±0.98％ vs 36.45％ ± 3.27％,P ＜ 0.05).The expression level of Dnmt1 mRNA in TCDD-exposure group was higher than that in control group on GD13.5(1.28±0.11 vs 1.01 ±0.10,P＜0.05) and on GD16.5(1.04 ±0.05 vs 0.81 ±0.01,P ＜0.01).The expression level of Dnmt3a mRNA in TCDD-exposure group was higher than that in control group on GD13.5 (1.15 ±0.17 vs 0.81 ±0.02,P ＜0.05)and on GD16.5 (1.11 ± 0.06 vs 0.96 ± 0.06,P ＜ 0.05).The expression level of Dnmt3b mRNA in TCDD-exposure group was higher than that in control group on GD14.5(0.97 ±0.06 vs 0.72 ±0.06,P ＜0.01).
Conclusions: It is supposed that complicated mechanisms are exist to regulate global DNA methylation levels in palatal tissue of fetal mice. The significant increased DNA methylation level on GD13.5 resulting from up-expression of Dnmt1 and Dnmt3a may be one of the epigenetic mechanisms which cause palate malformation in fetal mice induced by maternal exposure to TCDD.
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